Methods for the maturation of cardiomyocytes on amniotic fluid cell-derived ecm, cellular constructs, and uses for cardiotoxicity and proarrhythmic screening of drug compounds

ABSTRACT

Disclosed herein are methods of using a cell-derived extracellular matrix derived in-vitro from cells isolated from amniotic fluid (AFC-ECM) for the maturation of immature cardiomyocytes derived from human induced pluripotent stem cells (immature hiPSC-CMs) in culture forming mature cardiomyocytes. Also disclosed herein is a cell construct comprising a monolayer of these mature cardiomyocytes on an AFC-ECM useful for cardiotoxicity and/or proarrhythmic screening assays of drug compounds. Also disclosed herein are methods for determining the cardiotoxicity and/or proarrhythmic effect of a drug compound in vitro using such cell constructs.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional PatentApplication No. 62/808,690, filed Feb. 21, 2019, which is incorporatedby reference herein in its entirety

FIELD

The disclosure generally relates to the use of cellular constructs ofcardiomyocytes derived from human stem cells on cell-derivedextracellular matrices, methods of making the constructs, and methodsfor cardiotoxicity and proarrhythmic screening assays of drug compoundsusing the constructs.

BACKGROUND

Cardiotoxicity, or the perceived potential for cardiotoxicity, is aleading cause of toxicity related drug attrition during theinvestigation and selection of new drugs. Cardiac safety testing of newchemical entities that become lead drug candidates is a critical aspectof the drug discovery and development pipeline. A large number ofcardiac side effects of cardiac and non-cardiac drugs are caused by druginteraction with one or more cardiac ion channels. Cardiac ion channelsregulate cellular excitability, contractility and overall cardiacperformance, and alteration of cardiac ion channel function can lead tosudden cardiac death. This contributed to the release of drug candidatetesting guidelines from The International Conference on Harmonization(ICH).

The current preclinical drug candidate testing guidelines from the IHC(ICH S7A and S7B-Pharmacology Studies) rely on genetically modifiedheterologous cells and in vivo animal models. It has become increasinglyrecognized that these studies, such as hERG assay and QT prolongationstudies, do not accurately predict cardiotoxicity and proarrhythmia riskfor humans. Since 2005, cardiac safety of a drug compound has beendetermined almost exclusively by its effect or potential effect on theQT interval of the electrocardiogram (ECG) or the action potentialduration (APD), and the potential to lead to a life-threateningarrhythmia called Torsades de Pointes (TdP). However, QT prolongation isnot the ideal indicator for TdP as drugs that prolong the QT interval donot always cause TdP. It is now recognized that the QT prolongationparameter is only a surrogate marker for proarrhythmia. Data frompre-clinical and clinical trials have shown that there is no fixedrelationship between the magnitude of QT prolongation and the risk fordevelopment of fatal arrhythmias such as TdP.

Therefore, the US Food and Drug Administration (FDA) and otherstakeholders in drug discovery have called for an evolution inpre-clinical cardiotoxicity testing. The proposed new paradigm is calledthe Comprehensive In-Vitro Proarrhythmia Assay (CiPA). An integral partof the CiPA Initiative (http://cipaproject.org/) includes incorporationof data collected from human stem cell derived cardiomyocytes forcardiotoxicity and proarrhythmia assays. The overall goal of these newproposed guidelines is to provide a more accurate and comprehensivemechanistic based assessment of proarrhythmic potential that would moreaccurately assess the risk of new drugs. In review of the proposed CiPAInitiative's guidelines, the FDA has defined two advances that must bemade before human cardiomyocytes can be incorporated in the newinitiative. First, the growth and maturation state of human stem cellderived cardiomyocytes needs to be advanced to more closely resemble thestructure and function of adult human cardiomyocytes. Second, a reliablehigh throughput screening platform using these cells must be developed.

There are generally three types of systems currently utilized toevaluate the electrophysiology of cardiomyocytes in vitro: 1) Patchclamping systems; 2) micro-electrode array (MEA) systems; and 3) voltagesensitive dye (VSD) visualization methods. Manual patch clamping systemsare most commonly used in very early research studies to evaluate theelectrophysiology of individual cells. These devices, while providingaccurate and sensitive ionic current measurements, cannot be used for invitro cell systems that more closely mimic cell-cell interaction incardiac tissue. MEA systems include electrodes that are incorporatedinto cell culture wells allowing for the measurement of current acrossthe well. While allowing for high throughput analysis and ability tomeasure impedance in in vitro cell systems, these systems have lowspatial resolution, do not provide data on action potential shape, andhinder direct visualization of the cells and the ability to assess 3-Dculture systems that more closely mimic cardiac tissue architecture. VSDsystems are gaining interest as they address many of the shortcomings ofthe current technologies described above with features that include: 1)allowing for high throughput analysis; 2) providing high spatialresolution; 3) allowing visualization of impulse propagation across aculture dish; and 4) allowing for modification of the culture conditionsincluding addition of extracellular matrix leading to a more naturaltesting environment.

The use of cardiomyocytes derived from human stem cells, such as inducedpluripotent stem cells, have had limited success to date incardiotoxicity and proarrhythmic assay screening. Cardiomyocytes derivedfrom induced human pluripotent stem cells (hiPSC-CMs) are commerciallyavailable and can be purchased from several companies in cryopreservedvials that can be thawed and plated as monolayers. These hiPSC-CMs canbe made in large scale in vitro. Also, hiPSC-CMs can be obtained frompatient specific hiPSCs for a specific individual. However, there arehurdles that still need to be overcome to make the hiPSC-CMs ameaningful part of the new CiPA Initiative paradigm. 1) The maturationof hiPSC-CM structure and function must be advanced. Notably, the Kir2.1potassium channel is absent in currently available hiPSC-CMs and sodiumchannel expression is low. Currently available hiPSC-CMs are veryimmature functionally and structurally. The vast majority of currentlyused hiPSC-CM-based proarrhythmia screening assays rely on immature,fetal-like cells that do not resemble the structure of function of theadult cardiomyocyte. 2) Electrical pacing of hiPSC-CM monolayers in ahigh throughput electrophysiological screening platform is needed. Thevast majority of current hiPSC-CM-based proarrhythmia screens relysolely on field potential duration (MEA technology) or action potentialduration prolongation (VSD technology) as surrogate marker for TdPinduction. This is a limitation because not all drugs that prolong theaction potential (QT interval) will cause fatal TdP arrhythmias.Although certain maturation states of immature hiPSC-CMs have beenachieved using Matrigel™ ECM and bone-marrow cell-derived ECM inculture, use of these hiPSC-CMs in proarrhythmia screening assays didnot produce consistent results in the observation of arrhythmiaactivation patters consistent with what is known to occur in cases ofTdP in humans, especially between low risk and high risk drugs.

Thus, advanced materials and new methods are needed for the preciseassessment and screening of the cardiac safety liability of drugcompounds for accurate, reliable and efficient development of new drugcandidates.

SUMMARY

The present disclosure provides a solution to at least some of theaforementioned limitations and deficiencies in the art relating to theassessment and screening of the cardiac safety liability of drugcandidates. The solution is premised on the discovery of anextracellular matrix derived from cells isolated from amniotic fluid(AFC-ECM), which can be used for the maturation of human stem cellderived cardiomyocytes. These mature cardiomyocytes can then be used ina cellular construct with the AFC-ECM for cardiotoxicity andproarrhythmic testing of drugs.

In one aspect, disclosed is a method for the maturation of immaturecardiomyocytes derived from human induced pluripotent stem cells, themethod comprising: (a) providing immature cardiomyocytes derived fromhuman induced pluripotent stem cells (immature hiPSC-CMs); (b) providingan extracellular matrix derived in vitro from cells isolated fromamniotic fluid (AFC-ECM); (c) contacting the immature hiPSC-CMs with theAFC-ECM; and (d) culturing the immature hiPSC-CMs with the AFC-ECM in aculture media to induce maturation of the immature hiPSC-CMs, therebyforming mature cardiomyocytes; wherein the mature cardiomyocytes arecharacterized by rod shaped cells with distinct sarcomere structureresembling adult human cardiac tissue. In some embodiments the maturecardiomyocytes have a similar or the same morphology as illustrated inany one of FIGS. 6 to 14. In some embodiments, the immature hiPSC-CMsare plated on the AFC-ECM. In some embodiments, the maturecardiomyocytes form as a monolayer on the AFC-ECM, thereby forming acellular construct comprising a monolayer of mature cardiomyocytes onthe AFC-ECM, wherein the mature cardiomyocytes are aligned on theAFC-ECM. In some embodiments, the immature hiPSC-CMs do not expressinward-rectifier potassium channel Kir2.1. In some embodiments, theimmature hiPSC-CMs do not include rod shaped cells with distinctsarcomere structure. In some embodiments, the immature hiPSC-CMs can becharacterized by having less mitochondria than mature cardiomyocytes, byhaving disorganized myofilaments, by having a circular shape, and/or byhaving a single nucleus. In some embodiments, the mature cardiomyocytescan be further characterized by having greater amounts of mitochondriathan immature hiPSC-CMs, by having organized, compact myofilaments,and/or by having two nuclei (bi-nucleated).

In another aspect, disclosed is a cellular construct comprising amonolayer of mature cardiomyocytes on an extracellular matrix derivedfrom cells isolated in vitro from amniotic fluid (AFC-ECM), wherein themature cardiomyocytes are AFC-ECM cultured cardiomyocytes derived fromhuman induced pluripotent stem cells (hiPSC-CMs), and wherein the maturecardiomyocytes are characterized by rod shaped cells with distinctsarcomere structure resembling adult human cardiac tissue. In someembodiments the mature cardiomyocytes have a similar or the samemorphology as illustrated in any one of FIGS. 6 to 14. In some aspects,the mature cardiomyocytes include the inward-rectifier Kir2.1 potassiumchannel and/or express inward-rectifier potassium channel Kir2.1. Incertain aspects, the mature cardiomyocytes can be matured from immaturecardiomyocytes derived from human induced pluripotent stem cells(immature hiPSC-CMs) in culture on the AFC-ECM, wherein the maturecardiomyocytes are characterized by rod shaped cells with distinctsarcomere structure (striped appearance) resembling adult human cardiactissue. In some embodiments, the monolayer of mature cardiomyocytes isaligned on the AFC-ECM. In some embodiments, fiber tracks are present onthe construct. In some embodiments, the AFC-ECM comprises laminin,collagen alpha-1 (XVIII), basement membrane-specific heparan sulfateproteoglycan core protein, agrin, vimentin, and collagen alpha-2 (IV),and/or isoforms thereof. In some embodiments, the isoform of collagenalpha-1 (XVIII) is isoform 2. In some embodiments, the isoform of agrinis isoform 6. In some embodiments, the AFC-ECM further comprisesfibronectin and/or an isoform thereof. In some embodiments, the AFC-ECMdoes not contain decorin, perlecan, and/or collagen (III). In someembodiments, the immature hiPSC-CMs do not include rod shaped cells withdistinct sarcomere structure. In some embodiments, the immaturehiPSC-CMs can be characterized by having less mitochondria than maturecardiomyocytes, by having disorganized myofilaments, by having acircular shape, and/or by having a single nucleus. In some embodiments,the mature cardiomyocytes can be further characterized by having greateramounts of mitochondria than immature hiPSC-CMs, by having organized,compact myofilaments, and/or by having two nuclei (bi-nucleated).

In another aspect, disclosed is a method for making a cellular constructof mature cardiomyocytes on an extracellular matrix derived in vitrofrom cells isolated from amniotic fluid (AFC-ECM), the methodcomprising: (a) providing immature cardiomyocytes derived from humaninduced pluripotent stem cells (immature hiPSC-CMs); (b) providing anextracellular matrix derived in vitro from cells isolated from amnioticfluid (AFC-ECM); (c) plating the immature hiPSC-CMs on the AFC-ECM; and(d) culturing the plated immature hiPSC-CMs on the AFC-ECM in a culturemedia to induce maturation of the immature hiPSC-CMs into maturecardiomyocytes and to form a monolayer of the mature cardiomyocytes onthe AFC-ECM, thereby forming the cellular construct, wherein the maturecardiomyocytes are characterized by rod shaped cells with distinctsarcomere structure resembling adult human cardiac tissue. In someembodiments the mature cardiomyocytes have a similar or the samemorphology as illustrated in any one of FIGS. 6 to 14. In someembodiments, the monolayer of mature cardiomyocytes is aligned on theAFC-ECM. In some embodiments, fiber tracks are present on the cellularconstruct. In some embodiments, the immature hiPSC-CMs do not includerod shaped cells with distinct sarcomere structure. In some embodiments,the immature hiPSC-CMs can be characterized by having less mitochondriathan mature cardiomyocytes, by having disorganized myofilaments, byhaving a circular shape, and/or by having a single nucleus. In someembodiments, the mature cardiomyocytes can be further characterized byhaving greater amounts of mitochondria than immature hiPSC-CMs, byhaving organized, compact myofilaments, and/or by having two nuclei(bi-nucleated).

In another aspect, disclosed is a method for determining thecardiotoxicity and/or proarrhythmic effect of a drug compound in vitro,the method comprising contacting the drug compound with the maturecardiomyocytes of any one of the cellular constructs disclosedthroughout the specification, and observing for a change in theelectrophysiology of the mature cardiomyocytes to confirm whether thedrug compound has a cardiotoxic and/or proarrhythmic effect on themature cardiomyocytes. A change in the electrophysiology of the maturecardiomyocytes confirms that the drug compound has a cardiotoxic and/orproarrhythmic effect on the mature cardiomyocytes. The changes in theelectrophysiology of the mature cardiomyocytes can include, but are notlimited to, APD prolongation, APD prolongation plus rotors, and/orvarious types of arrhythmias, such as tachyarrhythmia (TA), quiescence(Q), delayed after depolarization (DAD), and/or early afterdepolarization (EAD). Observations can also include the cell viability,cell density, and/or morphology of the cells. In some embodiments, thechange in the electrophysiology of the mature cardiomyocytes isprolongation of action potential duration (APD). In some embodiments,the change in the electrophysiology of the mature cardiomyocytes isearly after depolarization (EAD). In some embodiments, the change in theelectrophysiology of the mature cardiomyocytes is delayed afterdepolarization (DAD). In some embodiments, the change in theelectrophysiology of the mature cardiomyocytes is action potentialduration prolongation (APD prolongation) plus rotors. In someembodiments, the change in the electrophysiology of the maturecardiomyocytes is an arrhythmia. In some aspects, the cellular constructcan be prepared by a process comprising: (a) providing immaturecardiomyocytes derived from human induced pluripotent stem cells(immature hiPSC-CMs); (b) providing an extracellular matrix derived invitro from cells isolated from amniotic fluid (AFC-ECM); (c) plating theimmature hiPSC-CMs on the AFC-ECM; and (d) culturing the plated immaturehiPSC-CMs on the AFC-ECM in a culture media to induce maturation of theimmature hiPSC-CMs into mature cardiomyocytes and to form a monolayer ofthe mature cardiomyocytes on the AFC-ECM, thereby forming a cellularconstruct, wherein the mature cardiomyocytes are characterized by rodshaped cells with distinct sarcomere structure resembling adult humancardiac tissue. In some embodiments the mature cardiomyocytes have asimilar or the same morphology as illustrated in any one of FIGS. 6 to14. In one embodiment, the immature hiPSC-CMs do not expressinward-rectifier potassium channel Kir2.1. In another embodiment, themonolayer of mature cardiomyocytes is aligned on the AFC-ECM. In anotherembodiment, fiber tracks are present on the cellular construct. In someembodiments, the immature hiPSC-CMs do not include rod shaped cells withdistinct sarcomere structure. In some embodiments, the immaturehiPSC-CMs can be characterized by having less mitochondria than maturecardiomyocytes, by having disorganized myofilaments, by having acircular shape, and/or by having a single nucleus. In some embodiments,the mature cardiomyocytes can be further characterized by having greateramounts of mitochondria than immature hiPSC-CMs, by having organized,compact myofilaments, and/or by having two nuclei (bi-nucleated).

Also, disclosed in the context of the present invention are thefollowing embodiments 1 to 20:

Embodiment 1 is a method for the maturation of immature cardiomyocytesderived from human induced pluripotent stem cells, the methodcomprising:(a) providing immature cardiomyocytes derived from human inducedpluripotent stem cells (immature hiPSC-CMs);(b) providing an extracellular matrix derived in vitro from cellsisolated from amniotic fluid (AFC-ECM);(c) contacting the immature hiPSC-CMs with the AFC-ECM; and(d) culturing the immature hiPSC-CMs with the AFC-ECM in a culture mediato induce maturation of the immature hiPSC-CMs, thereby forming maturecardiomyocytes;wherein the mature cardiomyocytes are characterized by rod shaped cellswith distinct sarcomere structure resembling adult human cardiac tissue.Embodiment 2 is the method of embodiment 1, wherein the immaturehiPSC-CMs are plated on the AFC-ECM.Embodiment 3 is the method of any one of embodiments 1 or 2, wherein themature cardiomyocytes form as a monolayer on the AFC-ECM, therebyforming a cellular construct comprising a monolayer of maturecardiomyocytes on the AFC-ECM, wherein the mature cardiomyocytes arealigned on the AFC-ECM.Embodiment 4 is the method of any one of embodiments 1 to 3, wherein theimmature hiPSC-CMs do not express inward-rectifier potassium channelKir2.1.Embodiment 5 is a cellular construct comprising a monolayer of maturecardiomyocytes on an extracellular matrix derived from cells isolated invitro from amniotic fluid (AFC-ECM), wherein the mature cardiomyocytesare AFC-ECM cultured cardiomyocytes derived from human inducedpluripotent stem cells (hiPSC-CMs), and wherein the maturecardiomyocytes are characterized by rod shaped cells with distinctsarcomere structure resembling adult human cardiac tissue.Embodiments 6 is the cellular construct of embodiment 5, wherein themonolayer of mature cardiomyocytes is aligned on the AFC-ECM.Embodiment 7 is the cellular construct of any one of embodiments 5 or 6,wherein fiber tracks are present on the construct.Embodiment 8 is the cellular construct of any one of embodiments 5 to 7,wherein the AFC-ECM comprises laminin, collagen alpha-1 (XVIII),basement membrane-specific heparan sulfate proteoglycan core protein,agrin, vimentin, and collagen alpha-2 (IV), and/or isoforms thereof.Embodiment 9 is the cellular construct of embodiment 8, wherein theisoform of collagen alpha-1 (XVIII) is isoform 2, and/or wherein theisoform of agrin is isoform 6.Embodiment 10 is the cellular construct of any one of embodiments 8 or9, wherein the AFC-ECM further comprises fibronectin and/or an isoformthereof.Embodiment 11 is the cellular construct of any one of embodiments 5 to10, wherein the AFC-ECM does not contain decorin, perlecan, and/orcollagen (III).Embodiment 12 is a method for making a cellular construct of maturecardiomyocytes on an extracellular matrix derived in vitro from cellsisolated from amniotic fluid (AFC-ECM), the method comprising:(a) providing immature cardiomyocytes derived from human inducedpluripotent stem cells (immature hiPSC-CMs),(b) providing an extracellular matrix derived in vitro from cellsisolated from amniotic fluid (AFC-ECM);(c) plating the immature hiPSC-CMs on the AFC-ECM;(d) culturing the plated immature hiPSC-CMs on the AFC-ECM in a culturemedia to induce maturation of the immature hiPSC-CMs into maturecardiomyocytes and to form a monolayer of the mature cardiomyocytes onthe AFC-ECM, thereby forming the cellular construct; wherein the maturecardiomyocytes are characterized by rod shaped cells with distinctsarcomere structure resembling adult human cardiac tissue.Embodiment 13 is the method of embodiment 12, wherein the monolayer ofmature cardiomyocytes is aligned on the AFC-ECM.Embodiment 14 is the method of any one of embodiments 12 or 13, whereinfiber tracks are present on the cellular construct.Embodiment 15 is a method for determining the cardiotoxicity and/orproarrhythmic effect of a drug compound in vitro, the method comprisingcontacting the drug compound with the mature cardiomyocytes of any oneof the cellular constructs of embodiments 5 to 11, and observing for achange in the electrophysiology of the mature cardiomyocytes to confirmwhether the drug compound has a cardiotoxic and/or proarrhythmic effecton the mature cardiomyocytes.Embodiment 16 is the method of embodiment 15, wherein the change in theelectrophysiology of the mature cardiomyocytes is prolongation of actionpotential duration (APD), and wherein prolongation of APD confirms thatthe drug compound has a cardiotoxic and/or proarrhythmic effect on themature cardiomyocytes.Embodiment 17 is the method of embodiment 15, wherein the change in theelectrophysiology of the mature cardiomyocytes is early afterdepolarization, and wherein early after depolarization (EAD) confirmsthat the drug compound has a cardiotoxic and/or proarrhythmic effect onthe mature cardiomyocytes.Embodiment 18 is the method of embodiment 15, wherein the change in theelectrophysiology of the mature cardiomyocytes is delayed afterdepolarization, and wherein delayed after depolarization (DAD) confirmsthat the drug compound has a cardiotoxic and/or proarrhythmic effect onthe mature cardiomyocytes.Embodiment 19 is the method of embodiment 15, wherein the change in theelectrophysiology of the mature cardiomyocytes is action potentialduration (APD) plus rotors, and wherein prolongation of APD plus rotorsconfirms that the drug compound has a cardiotoxic and/or proarrhythmiceffect on the mature cardiomyocytes.Embodiment 20 is the method of embodiment 15, wherein the change in theelectrophysiology of the mature cardiomyocytes is an arrhythmia, andwherein the arrhythmia confirms that the drug compound has a cardiotoxicand/or proarrhythmic effect on the mature cardiomyocytes.

The terms “about” or “approximately” are defined as being close to asunderstood by one of ordinary skill in the art, and in one non-limitingembodiment the terms are defined to be within 10%, preferably within 5%,more preferably within 1%, and most preferably within 0.5%.

The term “substantially” and its variations are defined to includeranges within 10%, within 5%, within 1%, or within 0.5%.

As used herein, the terms “% w/w” or “wt. %” refers to a weightpercentage of a component based on the total weight of material (e.g. acomposition) that includes the component. In a non-limiting example, 10grams of a component in 100 grams of a composition is 10% w/w of thecomponent in the total weight of composition. As used herein, the terms“% v/v” or “vol. %” refers to a volume percentage of a component basedon the total volume of material (e.g. a composition) that includes thecomponent. In a non-limiting example, 10 mL of a component in 100 mL ofa composition is 10% v/v of the component in the total volume ofcomposition. As used herein, the term “% w/v” refers to a weightpercentage of a component based on the total volume of material (e.g. acomposition) that includes the component. In a non-limiting example, 10grams of a component in 100 mL of a composition is 10% w/v of thecomponent in the total volume of composition. As used herein, the term“% v/w” refers to a volume percentage of a component based on the totalweight of material (e.g. a composition) that includes the component. Ina non-limiting example, 10 mL of a component in 100 grams of acomposition is 10% v/w of the component in the total weight of thecomposition.

The terms “inhibiting” or “reducing” or “preventing” or “avoiding” orany variation of these terms, when used in the claims and/or thespecification includes any measurable decrease or complete inhibition toachieve a desired result.

The term “effective,” as that term is used in the specification and/orclaims, means adequate to accomplish a desired, expected, or intendedresult.

The words “comprising” (and any form of comprising, such as “comprise”and “comprises”), “having” (and any form of having, such as “have” and“has”), “including” (and any form of including, such as “includes” and“include”) or “containing” (and any form of containing, such as“contains” and “contain”) are inclusive or open-ended and do not excludeadditional, unrecited elements or method steps.

The use of the word “a” or “an” when used in conjunction with the terms“comprising,” “having,” “including,” or “containing” (or any variationsof these words) may mean “one,” but it is also consistent with themeaning of “one or more,” “at least one,” and “one or more than one.”

The compositions and methods for their use can “comprise,” “consistessentially of,” or “consist of” any of the ingredients or stepsdisclosed throughout the specification. With respect to the transitionalphrase “consisting essentially of,” in one non-limiting aspect, a basicand novel characteristic of the cellular constructs disclosed herein istheir use in cardiotoxicity and/or proarrhythmic screening testing ofdrug compounds due to their ability to mature cardiomyocytes derivedfrom stem cells to a maturation state resembling that of mature nativeadult cardiomyocytes and native heart tissue.

It is contemplated that any embodiment discussed in this specificationcan be implemented with respect to any method or composition of theinvention, and vice versa. Furthermore, compositions of the inventioncan be used to achieve methods of the invention.

Other objects, features and advantages of the present invention willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and the specificexamples, while indicating specific embodiments of the invention, aregiven by way of illustration only, since various changes andmodifications within the spirit and scope of the invention will becomeapparent to those skilled in the art from this detailed description.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: A photomicrograph of a Brightfield Image of the amniotic fluidcell-derived ECM at 100× power using a 10× objective lens.

FIG. 2: An atomic force photomicrograph of 3 representative 40×40 umsections of the amniotic fluid cell-derived ECM and a bone marrowcell-derived ECM showing topography, adhesion, and stiffness.

FIG. 3: Scatter plots showing quantification of adhesion and stiffness(elastic modulus) of bone marrow- and amniotic fluid-cell-derived ECMs.Each point represents an independent point of measurement.

FIG. 4: A photomicrograph showing Day 0 and Day 2 culture of iPSCs onamniotic fluid cell-derived ECM and a bone marrow cell-derived ECM.

FIG. 5: A plot of growth curves of iPSCs cultured in the presence of theamniotic fluid cell-derived ECM and a bone marrow cell-derived ECM.

FIG. 6: A photomicrograph of mature cardiomyocytes on an AFC-ECM in asingle well—3.13 mm².

FIG. 7: A photomicrograph of mature cardiomyocytes on an AFC-ECM in asingle well—2.15 mm².

FIG. 8: A photomicrograph of mature cardiomyocytes on an AFC-ECM in asingle well—1.46 mm².

FIG. 9: A photomicrograph of mature cardiomyocytes on AFC-ECM in asingle well—0.54 mm².

FIG. 10: A photomicrograph of mature cardiomyocytes on AFC-ECM in asingle well—0.54 mm².

FIG. 11: A photomicrograph of mature cardiomyocytes on AFC-ECM in asingle well—0.54 mm².

FIG. 12: A photomicrograph of mature cardiomyocytes on AFC-ECM in asingle well—0.54 mm².

FIG. 13: A photomicrograph of mature cardiomyocytes on AFC-ECM in asingle well—0.13 mm².

FIG. 14: A photomicrograph of mature cardiomyocytes on AFC-ECM in asingle well—0.04 mm².

FIG. 15: A photomicrograph of cardiomyocytes on standard Matrigel™ ECMin a single well—3.13 mm².

FIG. 16: A photomicrograph of cardiomyocytes on standard Matrigel™ ECMin a single well—2.15 mm².

FIG. 17: A photomicrograph of cardiomyocytes on standard Matrigel™ ECMin a single well—1.46 mm².

FIG. 18: A photomicrograph of cardiomyocytes on standard Matrigel™ ECMin a single well—0.54 mm².

FIG. 19: A photomicrograph of cardiomyocytes on standard Matrigel™ ECMin a single well—0.13 mm².

FIG. 20: A photomicrograph of mature cardiomyocytes on a BM-ECM in asingle well—2.15 mm².

FIG. 21: A photomicrograph of mature cardiomyocytes on a BM-ECM in asingle well—1.46 mm².

FIG. 22: A schematic of the configuration of instruments to recordaction potential or calcium transient measurements using maturecardiomyocytes on AFC-ECM in multi-well plates.

FIG. 23: Recordings of spontaneous action potentials recorded fromcardiomyocytes cultured on Matrigel™ ECM, BM-ECM, and AFC-ECM, baselineand after drug E4031.

FIG. 24: Bar graph of number of Stable Rotors (TdP like arrhythmia) fromcardiomyocytes cultured on Matrigel™ ECM, BM-ECM, and AFC-ECM aftercontact with drug E4031.

FIG. 25: Recordings of spontaneous action potentials recorded frommature cardiomyocytes cultured on AFC-ECM for various drugs.

FIG. 26: Bar graph of total arrythmias recorded for all doses of each ofthe listed drugs.

FIG. 27: Bar graph of % of well with arrhythmia @ 10× the effectivetherapeutic plasma concentration (ETPC) of each of the listed drugs.

FIG. 28: Bar graph of the action potential triangulation (APD90-APD30)in time (ms) of each of the listed drugs.

FIG. 29: Bar graph of the action potential triangulation (APD90-APD30)in time (ms) of each of some of the listed drugs comparing cardiomyocyteperformance of cardiomyocytes on the AFC-ECM (SBS-AF Matrix) versus onMatrigel™ ECM.

FIG. 30: Bar graph of the maximum drug-induced action potentialtriangulation of the listed drugs comparing cardiomyocyte performance ofiCell® hiPSC-CMs from Cellular Dynamics (blank circles) versus Cor.4U®hiPSC-CMs from Ncardia (dark circles) at any concentration of the listeddrugs. Figure from Blinova et al, 2018, Cell Reports.

FIG. 31: Photomicrophraphs of hiPSC-CMs cultured on Matrigel™ ECM vs.AFC-ECM with immunofluorescentstaining for Troponin I, using DAPI tomark the nuclei.

FIG. 32: Photomicrophraphs of hiPSC-CMs cultured on Matrigel™ ECM vs.AFC-ECM with immunofluorescentstaining for α-actinin using DAPI to markthe nuclei.

FIG. 33: Photmicrographs of hiPSC-CMs cultured on Matrigel™ ECM vs.AFC-ECM with immunofluorescentstaining for cTnT and N Cadherin, usingDAPI to mark the nuclei.

FIG. 34: Photomicrographs of a single hiPSC-CM cell cultured onMatrigel™ ECM vs. AFC-ECM with immunofluorescentstaining for cTnT usingDAPI to mark the nuclei.

FIG. 35: Photomicrographs of a single hiPSC-CM cell cultured onMatrigel™ ECM vs. AFC-ECM with immunofluorescentstaining for α-actininusing DAPI to mark the nuclei.

FIG. 36: Graph of a comparison of the cellular circularity of the singlecells shown in FIG. 35.

FIG. 37: Photomicrographs of hiPSC-CMs cultured on Matrigel™ ECM vs.AFC-ECM with immunofluorescentstaining for cTnI expression, using DAPIto mark the nuclei.

FIG. 38: Western blotting of hiPSC-CMs on Matrigel™ ECM and AFC-ECM forcTnI expression and GAPDH.

FIG. 39: Graph of cTnI Expression relative to GAPDH of hiPSC-CMs onMatrigel™ ECM vs. AFC-ECM.

FIG. 40: Photomicrographs of hiPSC-CMs on Matrigel™ ECM vs. AFC-ECMstained for mitochondria with MitoTracker Red.

FIG. 41: Graph showing the MitoTraker™ Red fluorescenceintensity/cardiomyocyte for hiPSC-CMs on Matrigel™ ECM vs. AFC-ECM.

FIG. 42: Photomicrographs (transmitted light) of hiPSC-CMs cultured onMatrigel™ ECM and AFC-ECM that were coated on microelectrode array (MEA)plates.

DETAILED DESCRIPTION

Human stem cell derived cardiomyocytes, such as those derived from humaninduced pluripotent stem cells (hiPSC-CMs), that have been matured inculture on an extracellular matrix derived in vitro from cells isolatedfrom amniotic fluid (AFC-ECM) have demonstrated better, more consistentcardiotoxicity and proarrhythmia assay results than immature hiPSC-CMsor hiPSC-CMs matured on other ECMs, such as Matrigel™ ECM and bonemarrow cell-derived ECM. The hiPSC-CMs matured on an AFC-ECMsurprisingly have demonstrated a higher state of maturation thanhiPSC-CMs matured on other ECMs, such as Matrigel™ ECM and even on othernatural cell derived ECMs such as bone marrow cell-derived ECM, as shownby their cellular morphology and sarcomere structure that are found incardiomyocytes of normal adult human cardiac tissue. Furthermore, a morerobust expression of cTnI (cardiac troponin I) protein, as demonstratedby western blotting techniques, has been seen for hiPSC-CMs cultured onAFC-ECM than for hiPSC-CMs cultured on Matrigel™ ECM. Also, hiPSC-CMscultured on AFC-ECM have more mitochondria and mitochondria with morepolarized inner membrane potential than hiPSC-CMs cultured on Matrigel™ECM. Thus, the AFC-EMC can stimulate mitochondrial biogenesis,maturation, and function in the hiPSC-CMs. Normal adult or mature humancardiac human tissue is characterized by rod shaped cells with sarcomerestructure (striped appearance). The morphology and sarcomere structuresof the cardiomyocytes can be identified visually by microscopy(transmitted light or by immunofluorescent staining). As a non-limitingexample, the morphology and sarcomere structure of the cardiomyocytesmatured on AFC-ECM can be distinctly seen by the presence of rod shapedcells with a striped appearance identified by arrows in thephotomicrograph of FIG. 14. Additionally, the use of a siliconesubstrate such as PDMS were not needed to achieve these results.Surprisingly, a cellular construct comprising an AFC-ECM and a monolayerof mature cardiomyocytes that have been matured from immature hiPSC-CMsin culture on the AFC-ECM has shown to be useful, for example, byallowing consistent visualization of drug induced arrhythmias such asTorsades de Pointes (TdP) in high throughput in vitro screening assays.This “TdP in a dish” technology is a major advance over the reliance onfield potential duration (MEA technology) or action potential durationprolongation as surrogate markers for drug induced TdP. Thus, thecellular construct and methods disclosed herein go beyond the currentCiPA Initiative's guidelines by developing a more comprehensive andpredictive in vitro arrhythmia assay that allows for visualization ofarrhythmic events in an in vitro model, rather than simple polarizationand depolarization events that are indirect indicators of proarrhythmia.The cellular system and methods outlined in this disclosure allow forarrhythmic events to be propagated and visualized in an in vitro humancardiac monolayer model system. In some embodiments, the immaturehiPSC-CMs do not include rod shaped cells with distinct sarcomerestructure. In some embodiments, the immature hiPSC-CMs can becharacterized by having less mitochondria than mature cardiomyocytes, byhaving disorganized myofilaments, by having a circular shape, and/or byhaving a single nucleus. In some embodiments, the mature cardiomyocytescan be characterized by having rod shaped cells with distinct sarcomerestructure (striped appearance). In some embodiments, the maturecardiomyocytes can be further characterized by having greater amounts ofmitochondria than immature hiPSC-CMs, by having organized, compactmyofilaments, and/or by having two nuclei (bi-nucleated).

Disclosed herein are methods of using a cell-derived extracellularmatrix derived in-vitro from cells isolated from amniotic fluid(AFC-ECM) for the maturation of immature cardiomyocytes derived fromhuman induced pluripotent stem cells (immature hiPSC-CMs) in cultureforming mature cardiomyocytes (mature hiPSC-CMs). Also disclosed hereinis a cell construct comprising a monolayer of these maturecardiomyocytes on an AFC-ECM useful for cardiotoxicity and/orproarrhythmic screening assays of drug compounds. Also disclosed hereinare methods for determining the cardiotoxicity and/or proarrhythmiceffect of a drug compound in vitro using such cell constructs.

A. Amniotic Fluid Cell-Derived Extracellular Matrix (AFC-ECM)

Perinatal cells can be divided into three groups: cells from amnioticfluid; cells from the placenta; and cells from the umbilical cord.Amniotic fluid has several sources of cells including cells derived fromthe developing fetus sloughed from the fetal amnion membrane, skin, andalimentary, respiratory, and urogenital tracts. Placenta also hasseveral sources of cells including the membrane sheets (amnion andchorion), the villi, and the blood. Umbilical cord cells generally comefrom two sources, cord blood and Wharton's jelly. The cells from thesethree perinatal sources can include stem cells. The cells used toproduce the amniotic fluid cell-derived ECM of the invention areobtained from the amniotic fluid of a mammal including but not limitedto a human (Homo sapiens), murine, rabbit, cat, dog, pig, equine, orprimate. In preferred embodiments, the cells are from the amniotic fluidof a human. The amniotic fluid can be sourced from humans at full-termbirths (greater than about 37 weeks gestational age) or pre-term births(less than about 37 weeks gestational age). Pre-term births include latepre-term births (about 33 to about 37 weeks gestational age) moderatepre-term births (about 29 to about 33 weeks gestational age), andextreme pre-term births (about 23 to about 29 weeks gestational age).The amniotic fluid can be sourced from humans prior to birth at anygestational age where amniotic fluid is present, and can be combinedwith sources of amniotic fluid from births. Generally, prior to birth,amniotic fluid is collected by an amniocentesis procedure. In someembodiments the amniotic fluid is sourced from humans at full-termbirths, at pre-term births, at late pre-term births, at moderatepre-term births, at extreme pre-term births, or prior to birth, orcombinations thereof. In some embodiments, the amniotic fluid is sourcedprior to birth and is collected from about 10 weeks gestational age upto birth, or from about 10 weeks to about 23 weeks gestational age, orfrom about 10 weeks to about 16 weeks gestational age, or from about 12weeks gestational age up to birth, or from about 12 weeks to about 23weeks gestational age, or from about 12 weeks to about 16 weeksgestational age. In some embodiments, the amniotic fluid sourced priorto birth is collected by an amniocentesis procedure. The cells can beobtained and isolated from amniotic fluid by techniques known in theart, such as those disclosed in Murphy et. al., Amniotic Fluid StemCells, Perinatal Stem Cells, Second Ed. 2013.

Amniotic fluid is comprised of cells having the ability to differentiateinto cell types derived from all 3 embryonic germ layers (ectoderm,endoderm, mesoderm) spontaneously or as a result of treatment withspecific growth factors or combinations of growth factors known to oneof skill in the art. That is, a single cell has the capacity to beinduced to express genes which are specific to any of the three germlayers. Amniotic fluid also contains a mixture of different cell typesincluding cells derived from the developing fetus sloughed from thefetal amnion membrane, skin, and alimentary, respiratory, and urogenitaltracts. Because of the origin of the amniotic fluid and placentalmembranes, these cells can maintain highly multipotent differentiationpotential and comprise a cell population that contains cells of allthree germ layers. The amniotic fluid cells can comprise stem cells. Insome embodiments, the amniotic fluid cells are isolated stem cells. Insome embodiments, the amniotic fluid cells comprise stem cells havingthe ability to differentiate into cell types derived from all 3embryonic germ layers (ectoderm, endoderm, mesoderm) and/or multipotentstem cells, and/or pluripotent stem cells.

The amniotic fluid cell-derived ECM disclosed herein can comprisevarious proteins. The proteins of the ECM can be identified bytechniques known in the art and include mass spectroscopy andimmunohistochemical staining. The ECM can include, but is not limited tothe components listed in Table 2 (see Example 1 below) and any variants,derivatives, or isoforms thereof. The amniotic fluid-cell derived ECMcan include any combination of any of the components and any variants,derivatives, or isoforms thereof from Table 2. In some embodiments, acombination can comprise, consist essentially of, or consist of:laminin, collagen alpha-1 (XVIII), basement membrane-specific heparansulfate proteoglycan core protein, agrin, vimentin, and collagen alpha-2(IV), and/or isoforms thereof. In some embodiments, the isoform ofcollagen alpha-1 (XVIII) is isoform 2. In some embodiments, the isoformof agrin is isoform 6. In some embodiments, the cell-derived ECM furthercomprises, consist essentially of, or consists of fibronectin and/or anisoform thereof. In some embodiments, the amniotic fluid cell-derivedECM does not contain any one of or all of decorin, perlecan, andcollagen (III). Some noteworthy differences in proteins between theamniotic fluid cell-derived ECM of the present inventions and a bonemarrow cell-derived matrix are described in Table 1.

TABLE 1 Differences Between the Amniotic Fluid Cell-Derived ECM (AFC-Matrix) and a Bone Marrow Cell-Derived Matrix (BM-Matrix) Protein/Difference between AFC- Gene Code Matrix & BM-Matrix PhysiologicRelevance Laminin 5 sub-units are abundant in Laminin is known to AFCMatrix; Low expression support adhesion and in BM-Matrix expansion ofpluripotent cells. Collagen Abundant in AFC Matrix; Important for ocularXVIII Absent in BM-Matrix development Agrin Present in AFC Matrix;produced by motoneurons Absent in BM-Matrix to induce aggregation ofacetylcholine receptors Biglycan Abundant in BM-Matrix; Regulates boneand muscle Low expression in AFC development Matrix Collagen IOverexpressed in BM-Matrix Key to fibrillar proteins; relative to AFCMatrix highly abundant in bone Periostin Present in BM-Matrix; Regulatesmineralization; Absent in AFC Matrix Marker of noncardiomyocyte lineagecells in heart

The amniotic fluid cell-derived ECM can be produced by the followingprocess:

-   -   (a) isolating cells from amniotic fluid,    -   (b) seeding the isolated cells onto a cell culture container or        onto a cell culture container coated with a substrate,    -   (c) adding a culture media to the cell culture container, and    -   (d) culturing the cells, thereby producing a cell-derived ECM,        and    -   (e) optionally decellularizing the cell-derived ECM.

Any cell seeding density may be used which allows cells to form aconfluent monolayer immediately or after a period of time in culture. Insome embodiments, the seeding density is about 10 cells/cm²-about100,000 cells/cm², or about 100 cells/cm²-about 75,000 cells/cm², orabout 500 cells/cm²-about 50,000 cells/cm², or about 500 cells/cm²-about10,000 cells/cm², or about 500 cells/cm²-about 5,000 cells/cm², or about500 cells/cm²-about 2,500 cells/cm², or about 1,000 cells/cm²-about25,000 cells/cm², or about 2,000 cells/cm²-about 10,000 cells/cm², orabout 3,000 cells/cm²-about 5000 cells/cm².

Any type of container suitable for cultivation of cells can be used forthe present invention. Examples include, but are not limited to cellculture flasks, T-flasks, stirred flasks, spinner flasks, fermenters,and bioreactors. Rocking bottles, shaking flasks, tubes, and othercontainers are also suitable containers when placed on a rockingplatform or shaker. The cell culture container can be coated with asubstrate to allow for better cell adhesion. A non-limiting example of asuitable substrate for coating the cell container is fibronectin.

Various commercially available cell culture media, e.g., alpha MinimumEssential Media (α-MEM) culture media (Thermo Fisher Scientific, GrandIsland, N.Y.), are suitable for culturing amniotic fluid cells. Thecommercially available culture media can be modified by adding varioussupplemental substances to the media, e.g. sodium bicarbonate,L-glutamine, penicillin, streptomycin, Amphotericin B and/or serum. Theserum can be fetal bovine serum. The media can also be serum free.Additionally, substances such as L-ascorbic acid can be added to themedia or modified media to induce cell production of an ECM.

The initial culture media can be changed and/or replaced with anothermedia at various times during the culturing process. For example, theinitial media can be a “Complete Media” and then be replaced by an“Inducing Media” during the culturing process. A non-limiting example ofa “Complete Media” contains (α-MEM) plus 2 mM L-Glutamine plusantibiotic-antimycotic plus 15% Fetal Bovine Serum. A non-limitingexample of an “Inducing Media” contains the “Complete Media” plus 50 mML-Ascorbic Acid.

The culturing of the amniotic fluid cells can take place in an incubatorat 37° C., 5% CO₂, and 90% humidity. Culturing can take place undervarious environmental conditions including, but not limited to normoxic,i.e., 20-21% oxygen in the atmosphere, or hypoxic conditions.

Decellularizing the amniotic fluid cell-derived ECM of the amnioticfluid cells can include removing the viable amniotic fluid cells orrendering the amniotic fluid cells non-viable. The amniotic fluid cellscan be decellularized from the ECM by using methods known in the art andcan include, but are not limited to lysing the amniotic fluid cells andthen removing the lysed amniotic fluid cells by washing. Varioussubstances can be used to remove the amniotic fluid cells from the ECM.Non-limiting examples include an “Extraction Buffer” containing TRITONX-100 and ammonium hydroxide in PBS buffer. After the ECM has beendecellularized of amniotic fluid cells, the resulting ECM is therebyessentially cell-free or free of viable amniotic fluid cells. If feedercells are used, then the decellularizing methods also apply to anyviable feeder cells present on the ECM, thereby resulting in the ECMbeing essentially free or free of viable feeder cells. Thedecellularizing methods also apply to any viable cells present on theECM, thereby resulting in the ECM being essentially free or free of anyviable cells. Thus, a decellularized ECM means that the ECM isacellular, meaning that the ECM is free of any viable cells.

In some embodiments, the amniotic fluid cell-derived ECM (AFC-ECM) is athree-dimensional (3D) ECM.

The methods described supra also apply to producing cell-derived ECMsfrom other perinatal cells such as cells from the umbilical cordincluding the cord blood and Wharton's jelly; and cells from placentatissue including the membrane sheets (amnion and chorion), the villi andthe blood.

In one embodiment, a perinatal cell-derived ECM is produced by thefollowing process:

-   -   (a) isolating cells from an umbilical cord,    -   (b) seeding the isolated cells onto a cell culture container or        onto a cell culture container coated with a substrate,    -   (c) adding a culture media to the cell culture container, and    -   (d) culturing the cells, thereby producing a cell-derived ECM,        and    -   (e) optionally decellularizing the cell-derived ECM.

In some embodiments, the cells isolated from the umbilical cord are fromthe cord blood and/or the Wharton's jelly.

In another embodiment, a perinatal cell-derived ECM is produced by thefollowing process:

-   -   (a) isolating cells from placenta tissue,    -   (b) seeding the isolated cells onto a cell culture container or        onto a cell culture container coated with a substrate,    -   (c) adding a culture media to the cell culture container, and    -   (d) culturing the cells, thereby producing a cell-derived ECM,        and    -   (e) optionally decellularizing the cell-derived ECM.        In some embodiments, the cells isolated from the placenta tissue        are from the membrane sheets (amnion and/or chorion), the villi,        and/or the blood.

In one aspect, disclosed is a cell-derived extracellular matrix (ECM)derived in vitro from cells isolated from an umbilical cord. In someembodiments, the cells isolated from the umbilical cord are from thecord blood and/or the Wharton's jelly.

In another aspect, disclosed is a cell-derived extracellular matrix(ECM) derived in vitro from cells isolated from placenta tissue. In someembodiments, the cells isolated from the placenta tissue are from themembrane sheets (amnion and/or chorion), the villi, and/or the blood.

B. Cellular Constructs and Methods for the Maturation of ImmaturehiPSC-CMs

Disclosed herein is a method for the maturation of immaturecardiomyocytes derived from human induced pluripotent stem cells, themethod comprising: (a) providing immature cardiomyocytes derived fromhuman induced pluripotent stem cells (immature hiPSC-CMs); (b) providingan extracellular matrix derived in vitro from cells isolated fromamniotic fluid (AFC-ECM); (c) contacting the immature hiPSC-CMs with theAFC-ECM; and (d) culturing the immature hiPSC-CMs with the AFC-ECM in aculture media to induce maturation of the immature hiPSC-CMs, therebyforming mature cardiomyocytes (mature hiPSC-CMs); wherein the maturecardiomyocytes are characterized by rod shaped cells with distinctsarcomere structure resembling adult human cardiac tissue. Normal adulthuman cardiac human tissue is characterized by rod shaped cells withsarcomere structure (striped appearance). The morphology and sarcomerestructures of the cardiomyocytes can be identified visually bymicroscopy. As a non-limiting example, the morphology and sarcomerestructure of the cardiomyocytes matured on AFC-ECM can be distinctlyseen by the presence of rod shaped cells with a striped appearanceidentified by arrows in the photomicrograph of FIG. 14.

Also disclosed herein is a cellular construct comprising a monolayer ofmature cardiomyocytes on an extracellular matrix derived from cellsisolated in vitro from amniotic fluid (AFC-ECM), wherein the maturecardiomyocytes are AFC-ECM cultured cardiomyocytes derived from humaninduced pluripotent stem cells (hiPSC-CMs), and wherein the maturecardiomyocytes are characterized by rod shaped cells with distinctsarcomere structure resembling adult human cardiac tissue. In certainaspects, the mature cardiomyocytes can be matured from immaturecardiomyocytes derived from human induced pluripotent stem cells(immature hiPSC-CMs) in culture on the AFC-ECM, wherein the maturecardiomyocytes are characterized by rod shaped cells with distinctsarcomere structure resembling adult human cardiac tissue. In someaspects, the mature cardiomyocytes include the inward-rectifier Kir2.1potassium channel and/or express inward-rectifier potassium channelKir2.1. In some embodiments, the monolayer of mature cardiomyocytes isaligned on the AFC-ECM. In some embodiments, fiber tracks are present onthe construct. In some embodiments, the AFC-ECM comprises laminin,collagen alpha-1 (XVIII), basement membrane-specific heparan sulfateproteoglycan core protein, agrin, vimentin, and collagen alpha-2 (IV),and/or isoforms thereof. In some embodiments, the isoform of collagenalpha-1 (XVIII) is isoform 2, and/or wherein the isoform of agrin isisoform 6. In some embodiments, the AFC-ECM further comprisesfibronectin and/or an isoform thereof. In some embodiments, the AFC-ECMdoes not contain decorin, perlecan, and/or collagen (III). In someembodiments, the immature hiPSC-CMs do not include rod shaped cells withdistinct sarcomere structure. In some embodiments, the immaturehiPSC-CMs can be characterized by having less mitochondria than maturecardiomyocytes, by having disorganized myofilaments, by having acircular shape, and/or by having a single nucleus. In some embodiments,the mature cardiomyocytes can be further characterized by having greateramounts of mitochondria than immature hiPSC-CMs, by having organized,compact myofilaments, and/or by having two nuclei (bi-nucleated).

Also disclosed herein is a method for the preparation of a cellularconstruct comprising mature cardiomyocytes on an extracellular matrixderived in vitro from cells isolated from amniotic fluid (AFC-ECM), themethod comprising (a) providing immature cardiomyocytes derived fromhuman induced pluripotent stem cells (immature hiPSC-CMs), (b) providingan extracellular matrix derived in vitro from cells isolated fromamniotic fluid (AFC-ECM); (c) plating the immature hiPSC-CMs on theAFC-ECM; and (d) culturing the plated immature hiPSC-CMs on the AFC-ECMin a culture media to induce maturation of the immature hiPSC-CMs intomature cardiomyocytes and to form a monolayer of the maturecardiomyocytes on the AFC-ECM, thereby forming the cellular construct,wherein the mature cardiomyocytes are characterized by rod shaped cellswith distinct sarcomere structure resembling adult human cardiac tissue.In some embodiments, the monolayer of mature cardiomyocytes is alignedon the AFC-ECM. In some embodiments, fiber tracks are present on theconstruct.

Immature hiPSC-CMs can be obtained from commercial sources such asCellular Dynamics International-FUJI under the trade name iCell®, andTakara Bio under the trade name Cellartis®. iCell® Cardiomyocytes,iCell® Cardiomyocytes², and Cellartis® Cardiomyocytes are cryopreservedviable cardiomyocytes derived from human induced pluripotent stem cells(hiPSCs) available in vials. Immature hiPSC-CMs can also be generatedfrom patient specific hiPSCs in a laboratory setting for a specificindividual. In this case, differentiation of the hiPSCs can beaccomplished using the small molecule protocol to obtain beatingcardiomyocytes by day 8-10 using GSK3 inhibitor, RPMI/B27 minus insulin,Wnt Inhibitor and RPMI/B27 plus insulin at various days during the 7-daydifferentiation period. Suitable non-limiting examples of methods togenerate immature hiPSC-CMs are disclosed in US publication 2015/0329825herein incorporated by reference. In some embodiments, the immaturehiPSC-CMs do not express inward-rectifier potassium channel Kir2.1. Insome embodiments, the immature hiPSC-CMs do not include rod shaped cellswith distinct sarcomere structure. In some embodiments, the immaturehiPSC-CMs can be characterized by having less mitochondria than maturecardiomyocytes, by having disorganized myofilaments, by having acircular shape, and/or by having a single nucleus.

The AFC-ECM can be obtained using the production methods disclosedherein in this disclosure and can have the characteristics as describedin this disclosure. In some embodiments, the AFC-ECM is decellularizedprior to contact with the immature hiPSC-CMs.

The immature hiPSC-CMs can be in suspension when in contact with theAFC-ECM or the cells can be plated directly on the AFC-ECM which is inor on suitable cell culture containers or in multi-well plates.Non-limiting examples of suitable multi-well plates include 6-, 12-,24-, 48, 96- and 384-well plates. In some embodiments, the contactsurfaces of the cell culture containers or multi-well plates are coatedwith polydimethylsiloxane (PDMS) prior to the formation of the AFC-ECM.In some embodiments, the contact surfaces of the cell culture containersor multi-well plates are not coated with PDMS prior to the formation ofthe AFC-ECM. Any cell seeding density of immature hiPSC-CMs can be used.In some embodiments, a cell seeding density of immature hiPSC-CMs whichallows the cells to form a confluent monolayer immediately or after aperiod of time in culture is used. Cell density can be modified asdesired to improve monolayer formation. In multi-well plates, theimmature hiPSC-CM cells are placed in the center of each well.Non-limiting examples of cell seeding densities of immature hiPSC-CMs invarious multi-well plates are as follows: in 6-well plates, about200,000 cells can be plated per well; in 12-well plates, about 150,000cells can be plated per well; in 24-well plates about 175,000 cells canbe plated per well; in 48-well plates about 100,000 cells can be platedper well; in 96-well plates, about 50,000 cells can be plated per well;in 384-well plates, about 15,000 cells can be plated per well. In someembodiments, the cell seeding density of immature hiPSC-CMs is about50,000 cells per well in a 96-well plate. In some embodiments, the cellseeding density is about 200,000 cells per well in a 6-well plate. Toinduce maturation of the immature hiPSC-CMs, a suitable culture mediasuch as RPMI media or Media 199 media is added to the immature hiPSC-CMsin contact with the AFC-ECM, and the cells are cultured with the AFC-ECMusing standard cell culture techniques for a period of time, generally 7days, until the cells have matured into mature cardiomyocytes havingsimilar morphology as native adult cardiomyocytes. During the first 3 to4 days, the immature hiPCS-CMs are adhering, forming a continuousmonolayer, and beginning the maturation process. Surprisingly, theperiod of time for maturation of the cardiomyocytes can take 7 days orless, whereas generally a much longer period of time is typical withother substrates, e.g., up to 100 days. The morphology of these maturecardiomyocytes can be characterized by rod shaped cells with distinctsarcomere structure (striped appearance), the fundamental contractileunit of muscle, that can be visualized using conventional lightmicroscopy techniques. In some embodiments, the mature cardiomyocytescan be further characterized by having greater amounts of mitochondriathan immature hiPSC-CMs, by having organized, compact myofilaments,and/or by having two nuclei (bi-nucleated). Also, the AFC-ECM cannaturally produce fiber tracks that the mature cardiomyocytes follow.This produces a degree of anisotropy to the monolayer that more closelymimics the native heart. Thus, the cardiomyocytes can be naturallyaligned on the AFC-ECM following the alignment of the AFC-ECM that hasbeen laid down in a natural anisotropic configuration by the amnioticfluid cells during formation of the AFC-ECM. In various embodiments, theperiod of time for the immature hiPSC-CMs to mature in culture can be 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 40, 50, 60, 70, 80,90, 100, 110, or 120 days. Preferably, the period of time for thematuration of the cardiomyocytes is 14 days, or more preferably, 10days, or even more preferably 7 days. In some embodiments the period oftime for the immature hiPSC-CMs to mature in culture is 7 days. In someembodiments, the immature hiPSC-CMs are plated on the AFC-ECM. In someembodiments, the mature cardiomyocytes form as a confluent monolayer onthe AFC-ECM during culture, thereby forming a cellular constructcomprising a monolayer of mature cardiomyocytes on the AFC-ECM. In someembodiments, the mature cardiomyocytes are aligned on the AFC-ECM. Insome embodiments, the immature hiPSC-CMs are plated on the AFC-ECM inmulti-well plates. In some embodiments, the multi-well plates haveinserts of polydimethylsiloxane (PDMS). In some embodiment, themulti-well plates do not have inserts of PDMS.

C. Methods for Determining the Cardiotoxicity and/or ProarrhythmicEffect of a Drug Compound

Disclosed herein is a method for determining the cardiotoxicity and/orproarrhythmic effect of a drug compound in vitro, the method comprisingcontacting the drug compound with the mature cardiomyocytes of any oneof the cellular constructs disclosed throughout the specification, andobserving for one or more changes in the electrophysiology of the maturecardiomyocytes to confirm whether the drug compound has a cardiotoxicand/or proarrhythmic effect on the mature cardiomyocytes. One or morechanges in the electrophysiology of the mature cardiomyocytes indicatesand confirms that the drug compound has a cardiotoxic and/orproarrhythmic effect on the mature cardiomyocytes. The one or morechanges of the electrophysiology of the mature cardiomyocytes caninclude, but are not limited to, APD prolongation, APD prolongation plusrotors, and/or various types of arrhythmias, such as tachyarrhythmia(TA), quiescence (Q), delayed after depolarization (DAD), and/or earlyafter depolarization (EAD). In some embodiments, the change in theelectrophysiology of the mature cardiomyocytes is prolongation of actionpotential duration (APD). In some embodiments, the change in theelectrophysiology of the mature cardiomyocytes is early afterdepolarization (EAD). In some embodiments, the change in theelectrophysiology of the mature cardiomyocytes is delayed afterdepolarization (DAD). In some embodiments, the change in theelectrophysiology of the mature cardiomyocytes is action potentialduration (APD) plus rotors. In some embodiments, the change in theelectrophysiology of the mature cardiomyocytes is an arrhythmia. In someembodiments, the change in the electrophysiology of the maturecardiomyocytes is tachyarrhythmia (TA). In some embodiments, the changein the electrophysiology of the mature cardiomyocytes is quiescence (Q).Observations can also include the cell viability, cell density, and/ormorphology of the cells. In some aspects, the cellular construct can beprepared by a process comprising: (a) providing immature cardiomyocytesderived from human induced pluripotent stem cells (immature hiPSC-CMs);(b) providing an extracellular matrix derived in vitro from cellsisolated from amniotic fluid (AFC-ECM); (c) plating the immaturehiPSC-CMs on the AFC-ECM; and (d) culturing the plated immaturehiPSC-CMs on the AFC-ECM in a culture media to induce maturation of theimmature hiPSC-CMs into mature cardiomyocytes and to form a monolayer ofthe mature cardiomyocytes on the AFC-ECM, thereby forming a cellularconstruct, wherein the mature cardiomyocytes are characterized by rodshaped cells with distinct sarcomere structure resembling adult humancardiac tissue. In other aspects, the method can comprise contacting thedrug compound with any one of the cellular constructs disclosedthroughout the specification and observing for cardiotoxic and/orproarrhythmic events. In one particular instance, the method comprises:(a) providing immature cardiomyocytes derived from human inducedpluripotent stem cells (immature hiPSC-CMs); (b) providing anextracellular matrix derived in vitro from cells isolated from amnioticfluid (AFC-ECM); (c) plating the immature hiPSC-CMs on the AFC-ECM; (d)culturing the plated immature hiPSC-CMs on the AFC-ECM in a culturemedia to induce maturation of the immature hiPSC-CMs into maturecardiomyocytes and to form a monolayer of the mature cardiomyocytes onthe AFC-ECM, thereby forming a cellular construct; (e) contacting thedrug compound with the monolayer of the mature cardiomyocytes of theconstruct; and (f) observing for a change in the electrophysiology ofthe mature cardiomyocytes to confirm whether the drug compound has acardiotoxic and/or proarrhythmic effect on the mature cardiomyocytes;wherein the mature cardiomyocytes are characterized by rod shaped cellswith distinct sarcomere structure resembling adult human cardiac tissue.In some embodiments, the immature hiPSC-CMs are plated on the AFC-ECM ina multi-well plate. In some embodiments, the multi-well plates haveinserts of polydimethylsiloxane (PDMS). In some embodiment, themulti-well plates do not have inserts of PDMS. In some embodiments, theimmature hiPSC-CMs do not express inward-rectifier potassium channelKir2.1. In some embodiments, the monolayer of mature cardiomyocytes isaligned on the AFC-ECM. In some embodiments, fiber tracks are present onthe cellular construct.

The cardiotoxicity and/or proarrhythmia testing can be conducted usingany type of equipment suitable for measuring such activity. In someembodiments, the cellular constructs of monolayers of maturecardiomyocytes on AFC-ECM are prepared as described supra. After thematuration process (generally 7 days or less), the electrophysiology ofeach well is observed using a plate reader. A suitable voltage sensitiveor calcium sensitive fluorescent dye is loaded into each well.Non-limiting voltage sensitive dyes include FluoVolt™ dye commerciallyavailable from ThermoFisher. In some embodiments, the plate reader canrely on a suitable high spatiotemporal CCD camera combined with suitablelighting, such as light emitting diodes (LEDs), of the appropriatewavelength to excite each dye. Such high spatiotemporal CCD cameras arecommercially available from SciMeasure. In some embodiments, the cameraimage acquisition rate is greater than or equal to 150 frames persecond. In some embodiments, the camera and lens combination aredesigned such that it allows visualization of all the wells of themulti-well plate simultaneously with sufficient resolution to observeaction potential and calcium wave propagation. Each plate is centeredunder the camera system, lighting is switched on and camera acquisitionis initiated and electrophysiological activity is recorded. Experimentsare performed at about 37° C. After baseline readings are made, the drugto be tested is added to the wells and the effects are recorded.Spontaneous activity is recorded for a sufficient period of time toobtain images, e.g. at least 10 seconds. The images can be stored on acomputer. Images are analyzed and action potential duration, conductionvelocity, beat rate and activation patterns can be quantified usingimage analysis software. Visualization of electrical wave patterns isimportant to determine a drug compound's effect to cause potentiallyfatal arrhythmias, e.g., Torsades de Pointes (TdP). Thus, in addition toproviding information on a compound's effect on spontaneous actionpotential duration, the methods disclosed herein can also provideinformation on impulse conduction velocity and activation patterndepending on the type of equipment used.

D. Methods to Expand/Proliferate Mammalian Stem Cells

Also disclosed herein are methods of using a cell-derived extracellularmatrix derived in vitro from cells isolated from amniotic fluid(AFC-ECM) for the isolation, maintenance, and expansion/proliferation ofmammalian cells. In vitro cell culture is perhaps the most ubiquitous,important, and poorly understood aspect of all cell biology as well asthe developing fields of regenerative medicine and tissue engineering.Firstly, it allows for the observation of cell behavior so that variousaspects of cell function may be studied in detail. Secondly, it allowsfor increase in numbers of specific cell groups. For basic research, aswell as many clinical applications, it is necessary to achieve largequantities of relatively rare cells from small biological samples.In-vitro cell culture permits small numbers of cells to be expandedoutside the body to achieve more relevant numbers. Lastly, it permitsthe storage of cells for later use. By expanding cell numbers in-vitro,and freezing viable cells for later use, relatively small biologicalsamples can yield cells for multiple experiments over the span of days,months, or even years.

Despite the omnipresence of cell culture, the effects that in vitroculture has on the native characteristics of the cells is stillrelatively poorly understood. Many of the current practices have arisennot from deliberate thought, planning, and experimentation, but insteadfrom chance observations. Mammalian cell culture began in the early1900s when, in 1911, Alexis Carrel and Montrose Burrows first publishedan academic paper on the cultivation of mammalian tissues in vitro. Theywere studying the physiology and anatomy of tissues by cutting sectionsof mammalian tissues and placing them on microscope slides. They thennoticed that some cells migrated out of the tissue onto the slide. Theywent on to describe techniques for culturing cells in perpetuity. It nowappears that some of their observations may not have been valid, buttheir work paved the way for modern cell culture.

After the discovery of hematopoietic stem cells (HSCs), groups all overthe world were studying (HSCs). During their culture (in suspension), itwas observed that a sub-population of bone marrow cells stuck to thebottom of the plastic flasks and began to proliferate. These cells werelater recognized to be distinct from HSCs, and were eventually dubbedmesenchymal stem cells (MSCs). Because of this chance observation thatlead to the discovery of MSCs, plastic adherence is still widely used asa defining attribute of MSCs and many other mammalian cell types.

The practice of culturing cells on plastic substrates can be problematicbecause there is substantial evidence, that is now widely accepted inthe literature, demonstrating the critical role of the microenvironmentin regulating cell function. The microenvironment has been shown to helpdirect the differentiation of stem and progenitor cells, and regulatethe behavior of mature cell types.

When cells are removed from their native environment to be expandedin-vitro they can lose important cues from their surroundingextracellular matrix or microenvironment which relay importantinformation to the cells regarding the composition and state of theirsurroundings. Changes to a cell's microenvironment can have a profoundeffect on the behavior of those cells. The current standard forisolation and expansion of most adherent cells in vitro is to place thecells in culture vessels composed of polystyrene (plastic). Thepolystyrene may have been treated in some manner to facilitate cellattachment and growth but the surface is, in most cases, completelyforeign to the cell. In other cases, the surface may be coated withindividual matrix proteins (e.g. fibronectin or collagen) or somecombination of proteins. These simple substrates disregard thecomplexity of the native microenvironment as well as the critical roleof the microenvironment in normal cell function. The cell willimmediately begin to respond to this foreign environment in a mannerthat is much different than when the cell is in its native environment.

Five major approaches are currently employed to address this issue ofculturing cells on plastic substrates:

1. Ignore the problem.—Instead of trying to achieve a desired functionthat matches what would be expected in vivo, a multitude of cell typescan be tested in various media in order to find cells that will exhibita specific desired function without the appropriate matrix substrate.This approach is unsophisticated and often fails to produce desiredresults because of the complex interplay of variables and the breadth ofinteractions between cells and the extracellular matrix.

2. Identify key components.—Many academic laboratories and severalcompanies have taken the approach of considering the tissue from whichcells are isolated and looking for unique elements of that tissue thatmay be important for cell function. Cells are then cultured on simplesubstrates consisting of only one or a few matrix components. Thisapproach often fails because matrices are naturally very complexenvironments including over one-hundred different proteins in somecases. Cells respond just as strongly to signals they need and fail toreceive, as to signals they do not need and do receive.

3. Shotgun approach—The use of protein gels like MATRIGEL™ employs asort of shotgun approach. A gel is created that contains many differentmatrix proteins with the hopes that it will contain the necessarybinding motifs for many different cell types. This approach may fail byproviding cues that push cells in a particular direction or by failingto provide all the cues that cells are expecting.

4. Tissue-derived matrices—This is a biomimetic approach that typicallyinvolves isolating a tissue of interest from a genetically similaranimal, physically disrupting or chemically digesting the tissue toobtain a solution or uniform suspension, and then coating culturevessels with the deconstructed tissue. For example, someone who wishesto culture satellite cells, might collect muscle, homogenize the tissue,and then coat a culture vessel in homogenized muscle prior to seedingthe cells. This method often fails for a few reasons. Firstly, evenwithin a specific tissue type, the stem cell/progenitor cell niche, maybe distinct from the rest of the tissue. Simply homogenizing muscle doesnot guarantee that an appropriate niche is being created. Secondly, theniche consists of structural and physical cues, in addition tobiochemical cues. Even if many/most of the biochemical cues are presentin a tissue homogenate, the structure has been destroyed, and cells maysense very different mechanical cues. Lastly, manufacturability oftissue derived matrices is dependent on availability of tissues. Thisaffects the total amount of cell culture possible and contributes tolot-to-lot variability.

5. Cell-derived matrices—Cells in culture can be induced to secrete amatrix in their culture vessel. This matrix is the best approximationavailable of the in vivo niche, and can be manufactured in vitro. Cellscan be induced in vitro to elaborate a matrix and then the cells cansubsequently be eliminated from the matrix, for example by usingnon-ionizing detergent to retain structure and chemistry of the matrix.This approach has several key advantages. (1) The matrix structure canbe recreated and left undisturbed. (2) The matrix can be customizedbased on tissue/cell type of interest. (3) The matrix can be specific tostem and progenitor cell niche. (4) The matrix can be manufactured inlarge quantities.

With respect to cell-derived matrices, not all cell types can beefficiently isolated and expanded on any given cell-derived matrix. Infact, pluripotent stem cells (PSCs) appear to have much differentrequirements for a supportive growth substrate than do other types ofcells. It is known that the specific cell type used to produce a matrixwill have an effect on the composition of the matrix, and therefore, thereaction of various cell types to that matrix (see Marinkovic, M. etal., One size does not fit all: developing a cell-specific niche for invitro study of cell behavior. Matrix Biol. 54-55, 426-441 (2016)). Priorwork disclosed in U.S. Pat. No. 8,084,023 has described the productionand composition of an extracellular matrix produced by bone marrowstromal or mesenchymal stem cells (see also Chen, X. et al.,Extracellular Matrix Made by Bone Marrow Cells Facilitates Expansion ofMarrow-Derived Mesenchymal Progenitor Cells and Prevents TheirDifferentiation into Osteoblasts. Journal of Bone and Mineral Research22, 1943-1956 (2007) and Lai, Y. et al., Reconstitution ofmarrow-derived extracellular matrix ex vivo: a robust culture system forexpanding large-scale highly functional human mesenchymal stem cells.Stem cells and development 19, 1095-107 (2010)). This bone marrow cellderived matrix has been shown to support the expansion of other MSCs buthas not been effective for the attachment and growth of other types ofstem cells, specifically, induced pluripotent stem cells (iPSCs). iPSCshave exhibited an expanded potential to form cells and tissues from amuch broader category than MSCs. This represents a particularlyinteresting challenge, because the difficulty of growing a confluentmonolayer of iPSCs in standard culture conditions makes it impracticalto produce a cell-derived matrix from iPSCs. A major limitation ofprevious cell-derived matrices, is that in order to make atissue-specific matrix (e.g., bone marrow matrix from bone marrow MSCs,adipose matrix from adipose MSCs, or endothelial matrix from hUVECs), itis necessary that the target population of cells already be capable ofadhering to the starting substrate. A difficulty with iPSCs, embryonicstem cells (ES), and many other cell types is that they do not readilyadhere to simple substrates.

The present disclosure provides a solution to at least some of theaforementioned limitations and deficiencies in the art relating tocell-derived extracellular matrices (ECMs) to support the isolation,expansion and proliferation of pluripotent stem cells (PSCs), includingbut not limited to induced pluripotent stem cells (iPSCs) and embryonicstem cells (ES). The solution is premised on the use of an amnioticfluid cell-derived extracellular matrix. The use of uncommitted, readilyadherent, and highly proliferative perinatal cells found in amnioticfluid allows for the creation of an extracellular matrix (ECM) thatsurprisingly, supports adhesion, isolation, expansion, and proliferationof these PSCs. This technical achievement was not possible with thecell-derived ECMs of the prior art.

The function of mammalian cells is determined, largely, by theenvironment, e.g., an extracellular matrix, in which they reside. Theyreact to signals that are present in their environment (positivesignals) and also to signals that are required but are not present(negative signals). It is likely that uncommitted stem cells can producea matrix that contains niche motifs necessary to maintain stem cellviability and stemness, but lack many lineage specific signals that moremature cells may secrete which would push a stem cell toward aparticular fate. Without being bound by theory, it is suggested that aless mature cell, e.g., a perinatal cell or perinatal stem cell, mayproduce an ECM that is different from ECMs disclosed previously in theart, such as bone marrow stromal cell-derived ECMs, and may allow forbetter isolation and expansion/proliferation of stem cells with higherpotential than mesenchymal stem cells (MSCs), such as pluripotent stemcells (PSCs). Mass spectrometry demonstrated, that compared topreviously known cell-derived ECMs, the amniotic fluid cell-derived ECMof the invention contains matrix proteins found in all 3 germ layers andlacked specific proteins strongly associated with osteogenic lineages.Moreover, the ECM of the disclosure contain specific motifs, such aslaminin, that are known to facilitate pluripotent cell adhesion andexpansion.

Pluripotent stem cells (PSCs) can self-renew and differentiate into anyof the three germ layers: ectoderm, endoderm, and mesoderm, from whichall tissues and organs develop. Embryonic stem cells (ES) are currentlythe only known natural pluripotent stem cells. Induced pluripotent stem(iPSCs) cells also are PSCs. iPSCs are derived from cells generallytaken from adult tissue or adult cells, and reprogrammed to the level ofembryonic stem cells. Methods for producing iPSCs are known in the art.

Methods to expand/proliferate pluripotent stem cells (PSCs) includeobtaining PSCs and culturing them in the presence of the amniotic fluidcell-derived ECM of the invention. The PSCs can be iPSCs or ES. Anyseeding density may be used which allows cells to form a confluentmonolayer immediately or after a period of time in culture. In someembodiments, the seeding density is about 10 cells/cm²-about 100,000cells/cm², or about 100 cells/cm²-about 75,000 cells/cm², or about 500cells/cm²-about 50,000 cells/cm², or about 500 cells/cm²-about 10,000cells/cm², or about 500 cells/cm²-about 5,000 cells/cm², or about 500cells/cm²-about 2,500 cells/cm², or about 1,000 cells/cm²-about 25,000cells/cm², or about 2,000 cells/cm²-about 10,000 cells/cm², or about3,000 cells/cm²-about 5000 cells/cm².

In some embodiments the PSCs are maintained in an undifferentiated stateand maintain their stemness. Cell culture techniques suitable forproliferation of PSCs in culture are known in the art. Suitablecommercially available culture media for stem cell proliferationincludes, but is not limited to StemMACS™ iPS-Brew XF, available fromMiltenyl Biotec. In some embodiments, no Rock inhibitor is used. Oncecells begin to approach confluence (e.g., as determined by brightfieldmicroscopy), cells can be passaged manually, by cutting large coloniesinto smaller colonies and then re-plate those by physically lifting themoff the dish and placing them on a fresh plate of amniotic fluidcell-derived ECM. This procedure can be repeated indefinitely. In someembodiments disclosed is a method of proliferating pluripotent stemcells (PSCs) in culture, the method comprising culturing the PSCs in thepresence of a cell-derived extracellular matrix (ECM) in a culture mediathereby proliferating the PSCs, wherein the cell-derived ECM is derivedin-vitro from cells isolated from amniotic fluid.

The methods of expanding/proliferating PSCs described supra also applyto the use of expanding/proliferating PSCs in culture in the presence ofother perinatal cell-derived ECMs. In some embodiments, disclosed is amethod of proliferating pluripotent stem cells (PSCs) in culture, themethod comprising culturing the PSCs in the presence of a cell-derivedextracellular matrix (ECM) in a culture media thereby proliferating thePSCs, wherein the cell-derived ECM is derived in-vitro from cellsisolated from an umbilical cord or placenta tissue. In some embodiments,the cells isolated from the umbilical cord are from the cord bloodand/or the Wharton's jelly. In other embodiments, the cells isolatedfrom the placenta tissue are from the membrane sheets (amnion and/orchorion), the villi, and/or the blood.

EXAMPLES

The following examples are included to demonstrate certain non-limitingaspects of the invention. It should be appreciated by those of skill inthe art that the techniques disclosed in the examples that followrepresent techniques discovered by the applicants to function well inthe practice of the invention. However, those of skill in the artshould, in light of the present disclosure, appreciate that many changescan be made in the specific embodiments that are disclosed and stillobtain a like or similar result without departing from the spirit andscope of the invention.

Example 1—Production of an Amniotic Fluid Cell-Derived ECM (AFC-ECM)

Four amniotic fluid cell-derived ECMs (Matrix A, Matrix B, Matrix C, andMatrix D) were made using the following procedure: cells asepticallyisolated from amniotic fluid collected from full term birth (>37 weeksgestational age) from 4 donors were seeded onto fibronectin coatedtissue-culture treated flasks and cultured in Complete Media at 37° C.,5% CO₂ and 90% RH in an incubator. The Complete Media was alpha MinimumEssential Media (aMEM) plus 2 mM L-Glutamine plus antibiotic-antimycoticplus 15% Fetal Bovine Serum.

At day 3-4, one-half of the complete medium was aspirated from theflasks and replaced with one-half of new Complete Media. The flasks wereplaced back into the incubator at the same conditions as stated above.

At day 7-8, the Complete Media was aspirated from the culture flasks andwas replenished with Inducing Media. The flasks were placed back intothe incubator at the same conditions as stated above. The Inducing Mediawas Complete Media plus 50 mM L-Ascorbic Acid.

At day 10-11, the Inducing Media was aspirated from the culture flasksand the ECM which had formed inside the flasks was washed one time withphosphate buffered saline (PBS). Then the PBS was aspirated from theflasks. An Extraction Buffer was added to the flasks and incubated for7-10 minutes at RT to decellularize each ECM, then the Extraction Bufferwas aspirated from the flasks. The Extraction Buffer was PBS containing0.5% (v/v) TRITON-X100 and 20 mM ammonium hydroxide (NH₄OH).

Each of the decellularized ECMs in the flasks was washed three timeswith PBS followed by one wash with sterile water and then the sterilewater was aspirated from the flasks. The four decellularized ECMs in theflasks were allowed to dry at RT and then stored at 4° C.

A photomicrograph of a Brightfield Image of an amniotic fluidcell-derived ECM (Matrix B) is shown in FIG. 1 at 100× power using a 10×objective lens. An atomic force photomicrograph of 3 representative40×40 um sections of the amniotic fluid cell-derived ECM (Matrix B) anda bone marrow cell-derived ECM showing topography, adhesion, andstiffness is shown in FIG. 2. The bone marrow- and amnioticfluid-cell-derived ECMs are structurally and physically distinct.Quantification of adhesion and stiffness (elastic modulus) of bonemarrow- and amniotic fluid-cell-derived ECMs show bone marrow ECM is10-fold stiffer, and 3-fold less adhesive, relative to amniotic fluidECM as shown in the scatter plots in FIG. 3a (Adhesion) and FIG. 3b(Stiffness) where BM=bone marrow cell-derive ECM, AD=amniotic fluidcell-derived ECM (Matrix B). Each point represents an independent pointof measurement.

Example 2—Composition of Amniotic Fluid Cell-Derived ECM

The composition of the each of the amniotic fluid cell-derived ECMsproduced in Example 1 was determined by mass spectrometry. Thecomponents with their spectral count and molecular weight are listed inTable 2.

TABLE 2 Amniotic Fluid Cell-Derived ECM (AFC-ECM) Components TotalSpectra Count Matrix Matrix Matrix Matrix Protein MW A B C D Isoform 7of Fibronectin OS = Homo sapiens OX = 9606 269 kDa 817 13965 1143 794 GN= FN1 Isoform 3 of Fibronectin OS = Homo sapiens OX = 9606 259 kDa 80713836 1150 790 GN = FN1 Fibronectin OS = Homo sapiens OX = 9606 GN =FN1PE = 1 263 kDa 802 13851 1138 781 SV = 4 Isoform 14 of Fibronectin OS= Homo sapiens OX = 9606 249 kDa 770 12625 1103 763 GN = FN1 Isoform 10of Fibronectin OS = Homo sapiens OX = 9606 240 kDa 758 12506 1082 746 GN= FN1 Myosin-9 OS = Homo sapiens OX = 9606 GN = MYH9 227 kDa 564 3923295 340 PE = 1 SV = 4 SWISS-PROT: P60712 (Bos taurus) Actin,cytoplasmic1 42 kDa 293 2496 450 395 Vimentin OS = Homo sapiens OX =9606 GN = VIM PE = 1 54 kDa 263 1179 178 338 SV = 4 Neuroblastdifferentiation-associated protein AHNAK 629 kDa 250 803 17 184 OS =Homo sapiens OX = 9606 GN = AHNAK PE = 1 SV = 2 Histone H2B type 1-D OS= Homo sapiens OX = 9606 14 kDa 212 1121 176 187 GN = HIST1H2BD PE = 1SV = 2 Isoform 2 of Filamin-A OS = Homo sapiens OX = 9606 280 kDa 212342 12 143 GN = FLNA Basement membrane-specific heparan sulfate 469 kDa202 0 402 338 proteoglycan core protein OS = Homo sapiens OX = 9606 GN =HSPG2 PE = 1 SV = 4 Isoform 4 of Plectin OS = Homo sapiens OX = 9606 516kDa 185 337 10 83 GN = PLEC SWISS-PROT: P02769 (Bos taurus) Bovine serum69 kDa 122 112 53 72 albumin precursor Tubulin beta chain OS = Homosapiens OX = 9606 50 kDa 117 0 36 92 GN = TUBB PE = 1 SV = 2 Spectrinalpha chain, non-erythrocytic 1 OS = Homo 285 kDa 107 342 10 78 sapiensOX = 9606 GN = SPTAN1 PE = 1 SV = 3 Tubulin beta-4B chain OS = Homosapiens OX = 9606 50 kDa 107 0 34 84 GN = TUBB4B PE = 1 SV = 1 Isoform 3of Spectrin alpha chain, non-erythrocytic 1 282 kDa 106 345 0 77 OS =Homo sapiens GN = SPTAN1 Histone H4 OS = Homo sapiens OX = 9606 11 kDa100 1257 91 96 GN = HIST1H4A PE = 1 SV = 2 Tubulin beta-4A chain OS =Homo sapiens OX = 9606 50 kDa 99 0 31 76 GN = TUBB4A PE = 1 SV = 2Protein-glutamine gamma-glutamyltransferase 2 77 kDa 94 188 73 25 OS =Homo sapiens OX = 9606 GN = TGM2 PE = 1 SV = 2 SWISS-PROT:P00761|TRYP_PIG Trypsin - Sus scrofa 24 kDa 94 362 121 96 (Pig). Tubulinalpha-1B chain OS = Homo sapiens OX = 9606 50 kDa 93 295 37 86 GN =TUBA1B PE = 1 SV = 1 Isoform 2 of Clathrin heavy chain 1 OS = Homosapiens 188 kDa 92 97 1 15 OX = 9606 GN = CLTC Tubulin beta-2A chain OS= Homo sapiens OX = 9606 50 kDa 92 0 32 73 GN = TUBB2A PE = 1 SV = 1Elongation factor 1-alpha 1 OS = Homo sapiens OX = 9606 50 kDa 89 132 1470 GN = EEF1A1 PE = 1 SV = 1 Isoform 2 of Tubulin alpha-1A chain OS =Homo sapiens 46 kDa 88 305 34 82 OX = 9606 GN = TUBA1A Talin-1 OS = Homosapiens OX = 9606 GN = TLN1 PE = 1 270 kDa 88 109 4 41 SV = 3 Myosin-10OS = Homo sapiens GN = MYH10 PE = 1 SV = 3 229 kDa 84 417 56 46 Spectrinbeta chain, non-erythrocytic 1 OS = Homo 275 kDa 84 260 7 61 sapiens OX= 9606 GN = SPTBN1 PE = 1 SV = 2 Pyruvate kinase PKM OS = Homo sapiensOX = 9606 58 kDa 83 73 21 46 GN = PKM PE = 1 SV = 4 Tubulin alpha-1Cchain OS = Homo sapiens OX = 9606 50 kDa 83 0 0 78 GN = TUBA1C PE = 1 SV= 1 Major vault protein OS = Homo sapiens OX = 9606 99 kDa 82 360 68 108GN = MVP PE = 1 SV = 4 Actin, aortic smooth muscle OS = Homo sapiens 42kDa 78 0 126 77 OX = 9606 GN = ACTA2 PE = 1 SV = 1 Actin, gamma-entericsmooth muscle OS = Homo sapiens 42 kDa 76 313 122 73 OX = 9606 GN =ACTG2 PE = 1 SV = 1 Alpha-actinin-4 OS = Homo sapiens OX = 9606 105 kDa75 199 11 76 GN = ACTN4 PE = 1 SV = 2 Isoform 8 of Filamin-B OS = Homosapiens OX = 9606 282 kDa 75 123 2 94 GN = FLNB Tubulin alpha-4A chainOS = Homo sapiens OX = 9606 50 kDa 73 262 0 65 GN = TUBA4A PE = 1 SV = 1Glyceraldehyde-3-phosphate dehydrogenase OS = Homo 36 kDa 71 145 24 66sapiens OX = 9606 GN = GAPDH PE = 1 SV = 3 Cytoplasmic dynein 1 heavychain 1 OS = Homo sapiens 532 kDa 65 49 0 5 OX = 9606 GN = DYNC1H1 PE =1 SV = 5 Histone H2A.J OS = Homo sapiens OX = 9606 GN = H2AFJ 14 kDa 65216 62 79 PE = 1 SV = 1 Serpin H1 OS = Homo sapiens OX = 9606 GN =SERPINH1 46 kDa 64 731 128 97 PE = 1 SV = 2 Histone H2A type 2-C OS =Homo sapiens OX = 9606 14 kDa 63 0 61 82 GN = HIST2H2AC PE = 1 SV = 4Alpha-actinin-1 OS = Homo sapiens GN = ACTN1PE = 1 103 kDa 62 217 19 77SV = 2 Histone H2AX OS = Homo sapiens OX = 9606 15 kDa 61 0 69 75 GN =H2AFX PE = 1 SV = 2 Tubulin beta-3 chain OS = Homo sapiens OX = 9606 50kDa 58 194 28 53 GN = TUBB3 PE = 1 SV = 2 Endoplasmic reticulumchaperone BiP OS = Homo 72 kDa 57 0 35 46 sapiens OX = 9606 GN = HSPA5PE = 1 SV = 2 Myosin regulatory light chain 12A OS = Homo sapiens 20 kDa57 0 244 153 OX = 9606 GN = MYL12A PE = 1 SV = 1 Myosin regulatory lightchain 12B OS = Homo sapiens 20 kDa 57 0 240 149 OX = 9606 GN = MYL12B PE= 1 SV = 2 Elongation factor 2 OS = Homo sapiens OX = 9606 95 kDa 53 723 45 GN = EEF2 PE = 1 SV = 4 Filamin-C OS = Homo sapiens OX = 9606 GN =FLNC 291 kDa 53 62 3 63 PE = 1 SV = 3 Histone H3.1 OS = Homo sapiens OX= 9606 15 kDa 53 0 24 33 GN = HIST1H3A PE = 1 SV = 2 60 kDa heat shockprotein, mitochondrial OS = Homo 61 kDa 52 58 14 58 sapiens OX = 9606 GN= HSPD1 PE = 1 SV = 2 Tubulin beta-6 chain OS = Homo sapiens OX = 960650 kDa 52 0 23 52 GN = TUBB6 PE = 1 SV = 1 Isoform 4 of Collagenalpha-1(XII) chain OS = Homo 325 kDa 51 35 36 27 sapiens OX = 9606 GN =COL12A1 Nucleophosmin OS = Homo sapiens OX = 9606 33 kDa 48 103 15 43 GN= NPM1 PE = 1 SV = 2 Prelamin-A/C OS = Homo sapiens OX = 9606 GN = LMNA74 kDa 48 149 20 39 PE = 1 SV = 1 Heat shock protein HSP 90-beta OS =Homo sapiens 83 kDa 47 0 3 31 OX = 9606 GN = HSP90AB1 PE = 1 SV = 4 RasGTPase-activating-like protein IQGAP1 OS = Homo 189 kDa 47 0 11 36sapiens OX = 9606 GN = IQGAP1 PE = 1 SV = 1 Heat shock cognate 71 kDaprotein OS = Homo sapiens 71 kDa 46 130 28 56 OX = 9606 GN = HSPA8 PE =1 SV = 1 Annexin A2 OS = Homo sapiens OX = 9606 GN = ANXA2 39 kDa 43 1115 40 PE = 1 SV = 2 Isoform 2 of Collagen alpha-1(XVIII) chain OS = Homo154 kDa 42 575 77 61 sapiens OX = 9606 GN = COL18A1 Myosin regulatorylight polypeptide 9 OS = Homo sapiens 20 kDa 42 380 131 81 OX = 9606 GN= MYL9 PE = 1 SV = 4 Isoform 6 of Agrin OS = Homo sapiens OX = 9606 215kDa 41 168 42 70 GN = AGRN Histone H3.2 OS = Homo sapiens OX = 9606 15kDa 40 0 17 25 GN = HIST2H3A PE = 1 SV = 3 Isoform 1 of Core histonemacro-H2A.1 OS = Homo 39 kDa 40 450 43 49 sapiens OX = 9606 GN = H2AFYATP synthase subunit beta, mitochondrial OS = Homo 57 kDa 39 94 14 45sapiens OX = 9606 GN = ATP5F1B PE = 1 SV = 3 T-complex protein 1 subunitalpha OS = Homo sapiens 60 kDa 39 0 25 36 OX = 9606 GN = TCP1 PE = 1 SV= 1 Isoform 2 of Heat shock protein HSP 90-alpha 98 kDa 38 80 1 20 OS =Homo sapiens GN = HSP90AA1 Transitional endoplasmic reticulum ATPase OS= Homo 89 kDa 38 86 1 37 sapiens OX = 9606 GN = VCP PE = 1 SV = 4TREMBL: Q3KNV1; Q96GE1 Tax_Id = 9606 51 kDa 38 334 58 133 Gene_Symbol =KRT7 keratin 7 (Bos taurus) similar to alpha-2-macroglobulin isoform1164 kDa 37 14 9 13 Heterogeneous nuclear ribonucleoprotein U OS = Homo91 kDa 35 64 12 29 sapiens OX = 9606 GN = HNRNPU PE = 1 SV = 6 HistoneH3.3 OS = Homo sapiens OX = 9606 GN = H3F3A 15 kDa 35 0 17 43 PE = 1 SV= 2 Microtubule-associated protein 4 OS = Homo sapiens 121 kDa 35 73 826 OX = 9606 GN = MAP4 PE = 1 SV = 3 Beta-actin-like protein 2 OS = Homosapiens OX = 9606 42 kDa 34 147 42 31 GN = ACTBL2 PE = 1 SV = 2 Keratin,type I cytoskeletal 10 OS = Homo sapiens 59 kDa 34 464 32 37 OX = 9606GN = KRT10 PE = 1 SV = 6 Ribosome-binding protein 1 OS = Homo sapiens152 kDa 34 80 17 22 OX = 9606 GN = RRBP1 PE = 1 SV = 5Cytoskeleton-associated protein 4 OS = Homo sapiens 66 kDa 33 139 30 35OX = 9606 GN = CKAP4 PE = 1 SV = 2 DNA-dependent protein kinasecatalytic subunit 469 kDa 33 10 0 5 OS = Homo sapiens OX = 9606 GN =PRKDC PE = 1 SV = 3 Adenylyl cyclase-associated protein 1 OS = Homosapiens 52 kDa 32 33 2 37 OX = 9606 GN = CAP1 PE = 1 SV = 5 Keratin,type I cytoskeletal 9 OS = Homo sapiens 62 kDa 31 643 61 50 OX = 9606 GN= KRT9 PE = 1 SV = 3 Laminin subunit alpha-5 OS = Homo sapiens OX = 9606400 kDa 31 50 7 13 GN = LAMA5 PE = 1 SV = 8 (Bos taurus) similar tofibulin-1 C isoform 1 77 kDa 30 71 40 18 60S ribosomal protein L4 OS =Homo sapiens OX = 9606 48 kDa 30 75 26 25 GN = RPL4 PE = 1 SV = 5Alpha-enolase OS = Homo sapiens OX = 9606 GN = ENO1 47 kDa 30 61 3 26 PE= 1 SV = 2 ATP synthase subunit alpha, mitochondrial OS = Homo 60 kDa 3052 12 34 sapiens OX = 9606 GN = ATP5F1A PE = 1 SV = 1 Heterogeneousnuclear ribonucleoprotein M OS = Homo 78 kDa 30 114 12 30 sapiens OX =9606 GN = HNRNPM PE = 1 SV = 3 Histone H2A.V OS = Homo sapiens OX = 960614 kDa 30 131 23 34 GN = H2AFV PE = 1 SV = 3 Lysyl oxidase homolog 2 OS= Homo sapiens OX = 9606 87 kDa 30 320 94 52 GN = LOXL2 PE = 1 SV = 1MICOS complex subunit MIC60 OS = Homo sapiens 84 kDa 30 98 20 36 OX =9606 GN = IMMT PE = 1 SV = 1 SWISS-PROT: P12763 (Bos taurus) Alpha-2-HS-38 kDa 30 42 18 19 glycoprotein precursor Endoplasmin OS = Homo sapiensOX = 9606 92 kDa 29 71 3 22 GN = HSP90B1 PE = 1 SV = 1 TREMBL: Q0IIK2(Bos taurus) Transferrin 78 kDa 29 15 2 16 Laminin subunit beta-1 OS =Homo sapiens OX = 9606 198 kDa 28 30 4 11 GN = LAMB1 PE = 1 SV = 2T-complex protein 1 subunit beta OS = Homo sapiens 57 kDa 27 117 19 31OX = 9606 GN = CCT2 PE = 1 SV = 4 T-complex protein 1 subunit delta OS =Homo sapiens 58 kDa 27 73 19 26 OX = 9606 GN = CCT4 PE = 1 SV = 4Heterogeneous nuclear ribonucleoprotein K OS = Homo 51 kDa 26 28 3 26sapiens OX = 9606 GN = HNRNPK PE = 1 SV = 1 Bifunctionalglutamate/proline--tRNA ligase OS = Homo 171 kDa 25 20 0 1 sapiens OX =9606 GN = EPRS PE = 1 SV = 5 Isoform 1 of Vinculin OS = Homo sapiens OX= 9606 117 kDa 25 25 1 26 GN = VCL Isoform 2 of MICOS complex subunitMIC60 OS = Homo 83 kDa 25 99 17 34 sapiens OX = 9606 GN = IMMT T-complexprotein 1 subunit theta OS = Homo sapiens 60 kDa 25 100 21 30 OX = 9606GN = CCT8 PE = 1 SV = 4 Isoform LCRMP-4 of Dihydropyrimidinase-related74 kDa 24 11 4 40 protein 3 OS = Homo sapiens OX = 9606 GN = DPYSL3Thrombospondin-1 OS = Homo sapiens OX = 9606 129 kDa 24 103 3 1 GN =THBS1 PE = 1 SV = 2 Isoform Short of 14-3-3 protein beta/alpha OS = Homo28 kDa 23 18 2 14 sapiens OX = 9606 GN = YWHAB Isoform Smooth muscle ofMyosin light polypeptide 6 17 kDa 23 204 37 28 OS = Homo sapiens OX =9606 GN = MYL6 Keratin, type I cytoskeletal 18 OS = Homo sapiens 48 kDa23 376 99 120 OX = 9606 GN = KRT18 PE = 1 SV = 2 Keratin, type IIcytoskeletal 8 OS = Homo sapiens 54 kDa 23 1404 107 155 OX = 9606 GN =KRT8 PE = 1 SV = 7 Lamin-B1 OS = Homo sapiens OX = 9606 GN = LMNB1 66kDa 23 67 7 21 PE = 1 SV = 2 L-lactate dehydrogenase A chain OS = Homosapiens 37 kDa 23 18 2 10 GN = LDHA PE = 1 SV = 2 Profilin-1 OS = Homosapiens OX = 9606 GN = PFN1PE = 1 15 kDa 23 0 6 17 SV = 2 Proteindisulfide-isomerase OS = Homo sapiens OX = 9606 57 kDa 23 0 6 19 GN =P4HB PE = 1 SV = 3 T-complex protein 1 subunit gamma OS = Homo sapiens61 kDa 23 72 17 26 GN = CCT3 PE = 1 SV = 4 40S ribosomal protein S15 OS= Homo sapiens OX = 9606 17 kDa 22 0 26 38 GN = RPS15 PE = 1 SV = 2ATP-citrate synthase OS = Homo sapiens OX = 9606 121 kDa 22 20 2 18 GN =ACLY PE = 1 SV = 3 Isoform 2 of Transgelin-2 OS = Homo sapiens OX = 960624 kDa 22 47 1 20 GN = TAGLN2 Versican core protein OS = Homo sapiens OX= 9606 373 kDa 22 188 33 27 GN = VCAN PE = 1 SV = 3 60S acidic ribosomalprotein P0 OS = Homo sapiens 34 kDa 21 56 14 19 OX = 9606 GN = RPLP0 PE= 1 SV = 1 Histone H1.5 OS = Homo sapiens OX = 9606 23 kDa 21 53 11 20GN = HIST1H1B PE = 1 SV = 3 Importin subunit beta-1 OS = Homo sapiens OX= 9606 97 kDa 21 19 1 6 GN = KPNB1 PE = 1 SV = 2 Isoform 2 ofFructose-bisphosphate aldolase A 45 kDa 21 37 0 21 OS = Homo sapiens OX= 9606 GN = ALDOA Microtubule-associated protein 1B OS = Homo sapiens271 kDa 21 104 0 22 OX = 9606 GN = MAP1B PE = 1 SV = 2 TREMBL: Q3SX09(Bos taurus) similar to HBGprotein 22 kDa 21 26 11 9 5′-nucleotidase OS= Homo sapiens OX = 9606 GN = NT5E 63 kDa 20 129 21 17 PE = 1 SV = 1 60Sribosomal protein L3 OS = Homo sapiens OX = 9606 46 kDa 20 65 15 24 GN =RPL3 PE = 1 SV = 2 60S ribosomal protein L6 OS = Homo sapiens OX = 960633 kDa 20 0 19 25 GN = RPL6 PE = 1 SV = 3 ATP-dependent6-phosphofructokinase, platelet type 86 kDa 20 9 0 18 OS = Homo sapiensOX = 9606 GN = PFKP PE = 1 SV = 2 Collagen alpha-1(I) chain OS = Homosapiens OX = 9606 139 kDa 20 41 43 27 GN = COL1A1 PE = 1 SV = 5T-complex protein 1 subunit eta OS = Homo sapiens 59 kDa 20 72 15 22 OX= 9606 GN = CCT7 PE = 1 SV = 2 Trifunctional enzyme subunit alpha,mitochondrial 83 kDa 20 78 16 13 OS = Homo sapiens OX = 9606 GN = HADHAPE = 1 SV = 2 40S ribosomal protein S7 OS = Homo sapiens OX = 9606 22kDa 19 104 25 28 GN = RPS7 PE = 1 SV = 1 60S ribosomal protein L9 OS =Homo sapiens OX = 9606 22 kDa 19 0 14 21 GN = RPL9 PE = 1 SV = 1Actin-related protein 3 OS = Homo sapiens OX = 9606 47 kDa 19 0 16 18 GN= ACTR3 PE = 1 SV = 3 Annexin A5 OS = Homo sapiens OX = 9606 GN = ANXA536 kDa 19 0 0 15 PE = 1 SV = 2 Calpain-2 catalytic subunit OS = Homosapiens OX = 9606 80 kDa 19 4 0 8 GN = CAPN2 PE = 1 SV = 6 Heterogeneousnuclear ribonucleoprotein Al OS = Homo 39 kDa 19 55 10 23 sapiens OX =9606 GN = HNRNPA1 PE = 1 SV = 5 Integrin beta-1 OS = Homo sapiens OX =9606GN = ITGB1 88 kDa 19 16 4 18 PE = 1 SV = 2 Interleukinenhancer-binding factor 2 OS = Homo sapiens 43 kDa 19 0 4 15 OX = 9606GN = ILF2 PE = 1 SV = 2 Isoform 2 of Nidogen-2 OS = Homo sapiens OX =9606 141 kDa 19 55 0 3 GN = NID2 Matrin-3 OS = Homo sapiens OX = 9606 GN= MATR3 95 kDa 19 46 5 20 PE = 1 SV = 2 T-complex protein 1 subunit zetaOS = Homo sapiens 58 kDa 19 64 23 28 OX = 9606 GN = CCT6A PE = 1 SV = 340S ribosomal protein S2 OS = Homo sapiens OX = 9606 31 kDa 18 0 14 17GN = RPS2 PE = 1 SV = 2 40S ribosomal protein S3 OS = Homo sapiens OX =9606 27 kDa 18 82 19 20 GN = RPS3 PE = 1 SV = 2 Collagen alpha-2(IV)chain OS = Homo sapiens OX = 9606 168 kDa 18 209 44 33 GN = COL4A2 PE =1 SV = 4 Epiplakin OS = Homo sapiens OX = 9606 GN = EPPK1 556 kDa 18 221 39 PE = 1 SV = 3 Heat shock 70 kDa protein 1A OS = Homo sapiens 70 kDa18 31 0 23 OX = 9606 GN = HSPA1A PE = 1 SV = 1 Heterogeneous nuclearribonucleoproteins C1/C2 34 kDa 18 52 6 22 OS = Homo sapiens OX = 9606GN = HNRNPC PE = 1 SV = 4 Isoform 2 of Gelsolin OS = Homo sapiens OX =9606 81 kDa 18 87 36 24 GN = GSN Isoform 5 of Septin-9 OS = Homo sapiensOX = 9606 65 kDa 18 42 1 16 GN = SEPT9 Laminin subunit gamma-1 OS = Homosapiens OX = 9606 178 kDa 18 48 0 6 GN = LAMC1 PE = 1 SV = 3 NucleolinOS = Homo sapiens OX = 9606 GN = NCL PE = 1 77 kDa 18 58 8 16 SV = 3Voltage-dependent anion-selective channel protein 1 31 kDa 18 50 4 19 OS= Homo sapiens OX = 9606 GN = VDAC1 PE = 1 SV = 2 X-ray repaircross-complementing protein 6 OS = Homo 70 kDa 18 33 2 14 sapiens OX =9606 GN = XRCC6 PE = 1 SV = 2 Actin-related protein 2 OS = Homo sapiensOX = 9606 45 kDa 17 43 17 28 GN = ACTR2 PE = 1 SV = 1 Heat shock proteinbeta-1 OS = Homo sapiens OX = 9606 23 kDa 17 0 9 35 GN = HSPB1 PE = 1 SV= 2 Histone H1.4 OS = Homo sapiens OX = 9606 22 kDa 17 14 10 13 GN =HIST1H1E PE = 1 SV = 2 Isoform 2 of AP-2 complex subunit beta OS = Homo106 kDa 17 33 1 15 sapiens OX = 9606 GN = AP2B1 Isoform 2 of Calnexin OS= Homo sapiens OX = 9606 72 kDa 17 21 1 19 GN = CANX Isoform 2 ofDolichyl-diphosphooligosaccharid--protein 68 kDa 17 35 8 25glycosyltransferase subunit 2 OS = Homo sapiens OX = 9606 GN = RPN2Moesin OS = Homo sapiens OX = 9606 GN = MSN PE = 1 68 kDa 17 61 3 15 SV= 3 Protein disulfide-isomerase A3 OS = Homo sapiens 57 kDa 17 26 0 14OX = 9606 GN = PDIA3 PE = 1 SV = 4 Stress-70 protein, mitochondrial OS =Homo sapiens 74 kDa 17 33 0 10 OX = 9606 GN = HSPA9 PE = 1 SV = 2SWISS-PROT: P34955 (Bos taurus) Alpha-1- 46 kDa 17 66 19 16antiproteinase precursor T-complex protein 1 subunit epsilon OS = Homosapiens 60 kDa 17 110 18 22 OX = 9606 GN = CCT5 PE = 1 SV = 1 14-3-3protein zeta/delta OS = Homo sapiens OX = 9606 28 kDa 16 35 4 19 GN =YWHAZ PE = 1 SV = 1 40S ribosomal protein S4, X isoform OS = Homosapiens 30 kDa 16 77 20 20 OX = 9606 GN = RPS4X PE = 1 SV = 2 60Sribosomal protein L10 OS = Homo sapiens OX = 9606 25 kDa 16 27 7 12 GN =RPL10 PE = 1 SV = 4 Ezrin OS = Homo sapiens OX = 9606 GN = EZR PE = 1 69kDa 16 43 4 23 SV = 4 High mobility group protein HMG-I/HMG-Y OS = Homo12 kDa 16 52 7 11 sapiens OX = 9606 GN = HMGA1 PE = 1 SV = 3 Hypoxiaup-regulated protein 1 OS = Homo sapiens 111 kDa 16 37 0 18 OX = 9606 GN= HYOU1 PE = 1 SV = 1 Isoform 2 of Ubiquitin-like modifier-activatingenzyme 1 114 kDa 16 21 0 5 OS = Homo sapiens OX = 9606 GN = UBA1 Isoform3 of Heterogeneous nuclear ribonucleoprotein Q 63 kDa 16 37 5 20 OS =Homo sapiens OX = 9606 GN = SYNCRIP Nidogen-1 OS = Homo sapiens OX =9606 GN = NID1 136 kDa 16 56 1 7 PE = 1 SV = 3 14-3-3 protein epsilon OS= Homo sapiens OX = 9606 29 kDa 15 26 3 16 GN = YWHAE PE = 1 SV = 1 40Sribosomal protein S5 OS = Homo sapiens OX = 9606 23 kDa 15 0 10 12 GN =RPS5 PE = 1 SV = 4 60S ribosomal protein L5 OS = Homo sapiens OX = 960634 kDa 15 0 16 16 GN = RPL5 PE = 1 SV = 3 60S ribosomal protein L7 OS =Homo sapiens OX = 9606 29 kDa 15 61 14 14 GN = RPL7 PE = 1 SV = 1Caprin-1 OS = Homo sapiens OX = 9606 GN = CAPRIN1 78 kDa 15 18 0 14 PE =1 SV = 2 Catenin alpha-1 OS = Homo sapiens OX = 9606 100 kDa 15 13 0 12GN = CTNNA1 PE = 1 SV = 1 Eukaryotic initiation factor 4A-I OS = Homosapiens 46 kDa 15 28 2 8 OX = 9606 GN = EIF4A1 PE = 1 SV = 1Heterogeneous nuclear ribonucleoproteins A2/B1 37 kDa 15 58 6 20 OS =Homo sapiens OX = 9606 GN = HNRNPA2B1 PE = 1 SV = 2 Isoform 2 ofCoatomer subunit alpha OS = Homo sapiens 139 kDa 15 8 0 0 OX = 9606 GN =COPA Isoform 2 of Protein disulfide-isomerase A6 OS = Homo 54 kDa 15 230 15 sapiens OX = 9606 GN = PDIA6 Isoform 3 of Septin-2 OS = Homosapiens OX = 9606 43 kDa 15 55 6 16 GN = SEPT2 Staphylococcal nucleasedomain-containing protein 1 102 kDa 15 33 0 6 OS = Homo sapiens OX =9606 GN = SND1 PE = 1 SV = 1 Triosephosphate isomerase OS = Homo sapiensOX = 9606 31 kDa 15 15 0 11 GN = TPI1 PE = 1 SV = 3 UDP-glucose6-dehydrogenase OS = Homo sapiens 55 kDa 15 24 2 15 OX = 9606 GN = UGDHPE = 1 SV = 1 ADP/ATP translocase 2 OS = Homo sapiens OX = 9606 33 kDa14 27 9 17 GN = SLC25A5 PE = 1 SV = 7 ATP-dependent RNA helicase A OS =Homo sapiens 141 kDa 14 25 1 9 OX = 9606 GN = DHX9 PE = 1 SV = 4Cofilin-1 OS = Homo sapiens OX = 9606 GN = CFL1 PE = 1 19 kDa 14 0 4 11SV = 3 eIF-2-alpha kinase activator GCN1 OS = Homo sapiens 293 kDa 14 00 6 OX = 9606 GN = GCN1 PE = 1 SV = 6 Isoform 2 of Kinectin OS = Homosapiens OX = 9606 150 kDa 14 62 1 8 GN = KTN1 Isoform 2 of SpliceosomeRNA helicase DDX39B 51 kDa 14 25 1 12 OS = Homo sapiens OX = 9606 GN =DDX39B Isoleucine--tRNA ligase, cytoplasmic OS = Homo sapiens 145 kDa 140 0 0 OX = 9606 GN = IARS PE = 1 SV = 2 Keratin, type II cytoskeletal 2epidermal OS = Homo 65 kDa 14 123 28 21 sapiens OX = 9606 GN = KRT2 PE =1 SV = 2 Ribonuclease inhibitor OS = Homo sapiens OX = 9606 50 kDa 14 01 8 GN = RNH1 PE = 1 SV = 2 SWISS-PROT: P02070 (Bos taurus) Hemoglobinsubunit 16 kDa 14 0 0 0 beta SWISS-PROT: Q9DCV7 Tax_Id = 10090 51 kDa 140 14 18 Gene_Symbol = Krt7 Keratin, type II cytoskeletal 7Tubulointerstitial nephritis antigen-like OS = Homo 52 kDa 14 173 78 38sapiens OX = 9606 GN = TINAGL1 PE = 1 SV = 1 26S proteasome non-ATPaseregulatory subunit 2 100 kDa 13 18 0 10 OS = Homo sapiens OX = 9606 GN =PSMD2 PE = 1 SV = 3 40S ribosomal protein S8 OS = Homo sapiens OX = 960624 kDa 13 0 9 15 GN = RPS8 PE = 1 SV = 2 60S ribosomal protein L23 OS =Homo sapiens OX = 9606 15 kDa 13 0 14 10 GN = RPL23 PE = 1 SV = 1 60Sribosomal protein L7a OS = Homo sapiens OX = 9606 30 kDa 13 68 12 14 GN= RPL7A PE = 1 SV = 2 ADP/ATP translocase 3 OS = Homo sapiens OX = 960633 kDa 13 0 8 18 GN = SLC25A6 PE = 1 SV = 4 Arginine--tRNA ligase,cytoplasmic OS = Homo sapiens 75 kDa 13 33 0 5 OX = 9606 GN = RARS PE =1 SV = 2 Calpain small subunit 1 OS = Homo sapiens OX = 9606 28 kDa 13 00 22 GN = CAPNS1 PE = 1 SV = 1 Glycine--tRNA ligase OS = Homo sapiens OX= 9606 83 kDa 13 0 1 6 GN = GARS PE = 1 SV = 3 Isoform 2 of Extendedsynaptotagmin-1 OS = Homo 124 kDa 13 21 1 17 sapiens OX = 9606 GN =ESYT1 Isoform 2 of Nuclear mitotic apparatus protein 1 237 kDa 13 18 0 2OS = Homo sapiens OX = 9606 GN = NUMA1 Isoform 2 of Tropomyosin betachain OS = Homo sapiens 33 kDa 13 122 33 24 OX = 9606 GN = TPM2 Isoform7 of Interleukin enhancer-binding factor 3 96 kDa 13 31 0 13 OS = Homosapiens GN = ILF3 Isoform Beta-1 of DNA topoisomerase 2-beta OS = Homo183 kDa 13 97 3 3 sapiens OX = 9606 GN = TOP2B Lamin-B2 OS = Homosapiens OX = 9606 GN = LMNB2 70 kDa 13 53 8 19 PE = 1 SV = 4 L-lactatedehydrogenase B chain OS = Homo sapiens 37 kDa 13 0 0 6 OX = 9606 GN =LDHB PE = 1 SV = 2 Nucleoprotein TPR OS = Homo sapiens OX = 9606 267 kDa13 15 1 3 GN = TPR PE = 1 SV = 3 Receptor of activated protein C kinase1 OS = Homo 35 kDa 13 0 1 12 sapiens OX = 9606 GN = RACK1 PE = 1 SV = 3Serine protease HTRA1 OS = Homo sapiens OX = 9606 51 kDa 13 0 15 1 GN =HTRA1 PE = 1 SV = 1 Serine/threonine-protein phosphatase 2A 65 kDa 65kDa 13 0 0 10 regulatory subunit A alpha isoform OS = Homo sapiens OX =9606 GN = PPP2R1A PE = 1 SV = 4 X-ray repair cross-complementing protein5 OS = Homo 83 kDa 13 0 2 15 sapiens OX = 9606 GN = XRCC5 PE = 1 SV = 314-3-3 protein theta OS = Homo sapiens OX = 9606 28 kDa 12 0 2 12 GN =YWHAQ PE = 1 SV = 1 40S ribosomal protein S12 OS = Homo sapiens OX =9606 15 kDa 12 0 1 9 GN = RPS12 PE = 1 SV = 3 Cytoplasmic dynein 1 lightintermediate chain 2 54 kDa 12 12 3 19 OS = Homo sapiens OX = 9606 GN =DYNC1LI2 PE = 1 SV = 1 Glutathione S-transferase P OS = Homo sapiens OX= 9606 23 kDa 12 0 2 13 GN = GSTP1 PE = 1 SV = 2 Isoform 2 of Septin-7OS = Homo sapiens OX = 9606 51 kDa 12 31 0 12 GN = SEPT7 Isoform 3 ofUnconventional myosin-Ic OS = Homo 120 kDa 12 64 9 8 sapiens OX = 9606GN = MYO1C Peptidyl-prolyl cis-trans isomerase A OS = Homo sapiens 18kDa 12 36 3 11 OX = 9606 GN = PPIA PE = 1 SV = 2 Polyadenylate-bindingprotein 1 OS = Homo sapiens 71 kDa 12 22 0 15 OX = 9606 GN = PABPC1 PE =1 SV = 2 Rho-related GTP-binding protein RhoC OS = Homo 22 kDa 12 0 5 12sapiens OX = 9606 GN = RHOC PE = 1 SV = 1 Ribosomal L1 domain-containingprotein 1 OS = Homo 55 kDa 12 18 8 12 sapiens OX = 9606 GN = RSL1D1 PE =1 SV = 3 Transgelin OS = Homo sapiens OX = 9606 GN = TAGLN 23 kDa 12 4110 23 PE = 1 SV = 4 14-3-3 protein gamma OS = Homo sapiens OX = 9606 28kDa 11 0 1 14 GN = YWHAG PE = 1 SV = 2 26S proteasome regulatory subunit6A OS = Homo 47 kDa 11 0 0 17 sapiens OX = 9606 GN = PSMC3 PE = 1 SV = 140S ribosomal protein S17 OS = Homo sapiens OX = 9606 16 kDa 11 67 20 16GN = RPS17 PE = 1 SV = 2 60S ribosomal protein L10a OS = Homo sapiens 25kDa 11 0 11 17 OX = 9606 GN = RPL10A PE = 1 SV = 2 60S ribosomal proteinL12 OS = Homo sapiens OX = 9606 18 kDa 11 36 10 9 GN = RPL12 PE = 1 SV =1 ADP-ribosylation factor 4 OS = Homo sapiens OX = 9606 21 kDa 11 0 8 14GN = ARF4 PE = 1 SV = 3 Annexin A6 OS = Homo sapiens OX = 9606 GN =ANXA6 76 kDa 11 15 0 9 PE = 1 SV = 3 ATP synthase subunit O,mitochondrial OS = Homo 23 kDa 11 0 5 10 sapiens OX = 9606 GN = ATP5POPE = 1 SV = 1 Core histone macro-H2A.2 OS = Homo sapiens OX = 9606 40kDa 11 226 28 21 GN = H2AFY2 PE = 1 SV = 3Dolichyl-diphosphooligosaccharide--protein 69 kDa 11 43 7 18glycosyltransferase subunit 1 OS = Homo sapiens OX = 9606 GN = RPN1 PE =1 SV = 1 Elongation factor 1-gamma OS = Homo sapiens OX = 9606 50 kDa 1129 0 8 GN = EEF1G PE = 1 SV = 3 Guanine nucleotide-binding protein G(i)subunit alpha-2 40 kDa 11 45 6 8 OS = Homo sapiens OX = 9606 GN = GNAI2PE = 1 SV = 3 Heterogeneous nuclear ribonucleoprotein R OS = Homo 71 kDa11 17 4 12 sapiens OX = 9606 GN = HNRNPR PE = 1 SV = 1 Isoform 2 ofProbable ATP-dependent RNA helicase 72 kDa 11 17 0 9 DDX17 OS = Homosapiens OX = 9606 GN = DDX17 Isoform 3 of Tropomyosin alpha-1 chain OS =Homo 33 kDa 11 181 46 37 sapiens OX = 9606 GN = TPM1 Isoform 4 ofCaldesmon OS = Homo sapiens OX = 9606 63 kDa 11 51 10 32 GN = CALD1Leucine--tRNA ligase, cytoplasmic OS = Homo sapiens 134 kDa 11 3 0 0 GN= LARS PE = 1 SV = 2 Phosphoglycerate kinase 1 OS = Homo sapiens OX =9606 45 kDa 11 10 0 7 GN = PGK1 PE = 1 SV = 3 Polypyrimidinetract-binding protein 1 OS = Homo 57 kDa 11 33 1 20 sapiens OX = 9606 GN= PTBP1 PE = 1 SV = 1 Rab GDP dissociation inhibitor beta OS = Homosapiens 51 kDa 11 17 0 10 OX = 9606 GN = GDI2 PE = 1 SV = 2 Reticulon-4OS = Homo sapiens GN = RTN4 PE = 1 SV = 2 130 kDa 11 15 0 9 Serinehydroxymethyltransferase, mitochondrial 56 kDa 11 6 0 3 OS = Homosapiens GN = SHMT2 PE = 1 SV = 3 Serine/threonine-protein phosphatasePP1-alpha catalytic 38 kDa 11 23 3 10 subunit OS = Homo sapiens OX =9606 GN = PPP1CA PE = 1 SV = 1 Splicing factor, proline- andglutamine-rich OS = Homo 76 kDa 11 43 5 18 sapiens OX = 9606 GN = SFPQPE = 1 SV = 2 SWISS-PROT: Q3MHN5 (Bos taurus) Vitamin D- 53 kDa 11 15 24 binding protein precursor Tenascin OS = Homo sapiens GN = TNC PE = 1SV = 3 241 kDa 11 409 60 72 Threonine--tRNA ligase, cytoplasmic OS =Homo sapiens 83 kDa 11 10 0 3 OX = 9606 GN = TARS PE = 1 SV = 3Tropomyosin alpha-4 chain OS = Homo sapiens OX = 9606 29 kDa 11 89 21 20GN = TPM4 PE = 1 SV = 3 14-3-3 protein eta OS = Homo sapiens OX = 960628 kDa 10 0 1 10 GN = YWHAH PE = 1 SV = 4 26S proteasome regulatorysubunit 6B OS = Homo sapiens 47 kDa 10 13 3 10 OX = 9606 GN = PSMC4 PE =1 SV = 2 60S ribosomal protein L18 OS = Homo sapiens 19 kDa 10 26 7 9 GN= RPL18 PE = 1 SV = 1 Actin-related protein 2/3 complex subunit 2 OS =Homo 34 kDa 10 53 14 11 sapiens OX = 9606 GN = ARPC2 PE = 1 SV = 1A-kinase anchor protein 12 OS = Homo sapiens OX = 9606 191 kDa 10 46 0 5GN = AKAP12 PE = 1 SV = 4 Asparagine--tRNA ligase, cytoplasmic OS = Homo63 kDa 10 10 0 1 sapiens OX = 9606 GN = NARS PE = 1 SV = 1Aspartate--tRNA ligase, cytoplasmic OS = Homo sapiens 57 kDa 10 22 3 9OX = 9606 GN = DARS PE = 1 SV = 2Dolichyl-diphosphooligosaccharide--protein 51 kDa 10 13 6 17glycosyltransferase 48 kDa subunit OS = Homo sapiens OX = 9606 GN =DDOST PE = 1 SV = 4 Erlin-2 OS = Homo sapiens OX = 9606 GN = ERLIN2 PE =1 38 kDa 10 39 4 11 SV = 1 Fatty acid synthase OS = Homo sapiens OX =9606 273 kDa 10 0 0 2 GN = FASN PE = 1 SV = 3 Heterogeneous nuclearribonucleoprotein A3 OS = Homo 40 kDa 10 25 7 8 sapiens OX = 9606 GN =HNRNPA3 PE = 1 SV = 2 Isoform 3 of Exportin-2 OS = Homo sapiens OX =9606 108 kDa 10 2 2 3 GN = CSE1L Isoform B of AP-1 complex subunitbeta-1 OS = Homo 104 kDa 10 0 0 11 sapiens OX = 9606 GN = AP1B1Leucine-rich PPR motif-containing protein, 158 kDa 10 5 0 0mitochondrial OS = Homo sapiens OX = 9606 GN = LRPPRC PE = 1 SV = 3Non-POU domain-containing octamer-binding protein 54 kDa 10 44 3 20 OS =Homo sapiens OX = 9606 GN = NONO PE = 1 SV = 4 Nucleolar protein 56 OS =Homo sapiens OX = 9606 66 kDa 10 18 1 2 GN = NOP56 PE = 1 SV = 4Peroxidasin homolog OS = Homo sapiens OX = 9606 165 kDa 10 146 36 28 GN= PXDN PE = 1 SV = 2 Stomatin-like protein 2, mitochondrial OS = Homosapiens 39 kDa 10 18 3 10 OX = 9606 GN = STOML2 PE = 1 SV = 1SWISS-PROT: P01966 (Bos taurus) Hemoglobin subunit 15 kDa 10 34 12 10alpha SWISS-PROT: Q3SZ57 (Bos taurus) Alpha-fetoprotein 69 kDa 10 11 6 7precursor Transforming protein RhoA OS = Homo sapiens 22 kDa 10 0 4 10OX = 9606 GN = RHOA PE = 1 SV = 1 40S ribosomal protein SA OS = Homosapiens OX = 9606 33 kDa 9 0 1 10 GN = RPSA PE = 1 SV = 1 60S ribosomalprotein L13 OS = Homo sapiens OX = 9606 24 kDa 9 32 12 12 GN = RPL13 PE= 1 SV = 4 60S ribosomal protein L8 OS = Homo sapiens OX = 9606 28 kDa 90 7 9 GN = RPL8 PE = 1 SV = 2 Aldehyde dehydrogenase X, mitochondrial OS= Homo 57 kDa 9 14 4 11 sapiens OX = 9606 GN = ALDH1B1 PE = 1 SV = 3ATP-dependent RNA helicase DDX3X OS = Homo 73 kDa 9 18 5 11 sapiens OX =9606 GN = DDX3X PE = 1 SV = 3 Calreticulin OS = Homo sapiens OX = 9606GN = CALR 48 kDa 9 44 4 11 PE = 1 SV = 1 D-3-phosphoglyceratedehydrogenase OS = Homo sapiens 57 kDa 9 0 0 2 OX = 9606 GN = PHGDH PE =1 SV = 4 F-actin-capping protein subunit alpha-1 OS = Homo 33 kDa 9 0 1815 sapiens OX = 9606 GN = CAPZA1 PE = 1 SV = 3 Galectin-1 OS = Homosapiens OX = 9606 GN = LGALS1 15 kDa 9 0 3 6 PE = 1 SV = 2 GTP-bindingnuclear protein Ran OS = Homo sapiens 24 kDa 9 0 6 11 OX = 9606 GN = RANPE = 1 SV = 3 Heterochromatin protein 1-binding protein 3 OS = Homo 61kDa 9 24 4 7 sapiens OX = 9606 GN = HP1BP3 PE = 1 SV = 1 Isoform 1 ofVoltage-dependent anion-selective channel 33 kDa 9 19 1 5 protein 2 OS =Homo sapiens OX = 9606 GN = VDAC2 Isoform 2 of Eukaryotic translationinitiation factor 3 163 kDa 9 10 0 11 subunit A OS = Homo sapiens OX =9606 GN = EIF3A Isoform 2 of Glutamine--tRNA ligase OS = Homo sapiens 87kDa 9 16 0 4 OX = 9606 GN = QARS Isoform 2 of Tropomyosin alpha-3 chainOS = Homo 29 kDa 9 99 16 17 sapiens OX = 9606 GN = TPM3 Isoform 3 ofProtein AHNAK2 OS = Homo sapiens 606 kDa 9 0 0 1 OX = 9606 GN = AHNAK2Isoform D of Eukaryotic translation initiation factor 4 159 kDa 9 7 0 0gamma 1 OS = Homo sapiens OX = 9606 GN = EIF4G1 KH domain-containing,RNA-binding, signal 48 kDa 9 24 5 9 transduction-associated protein 1 OS= Homo sapiens OX = 9606 GN = KHDRBS1 PE = 1 SV = 1 Methionine--tRNAligase, cytoplasmic OS = Homo 101 kDa 9 1 0 0 sapiens OX = 9606 GN =MARS PE = 1 SV = 2 Myeloid-associated differentiation marker OS = Homo35 kDa 9 0 16 12 sapiens OX = 9606 GN = MYADM PE = 1 SV = 2 Neutralalpha-glucosidase AB OS = Homo sapiens 107 kDa 9 21 0 13 OX = 9606 GN =GANAB PE = 1 SV = 3 Peroxiredoxin-5, mitochondrial OS = Homo sapiens 22kDa 9 6 0 5 OX = 9606 GN = PRDX5 PE = 1 SV = 4 Serine--tRNA ligase,cytoplasmic OS = Homo sapiens 59 kDa 9 0 0 9 OX = 9606 GN = SARS PE = 1SV = 3 Tryptophan--tRNA ligase, cytoplasmic OS = Homo 53 kDa 9 6 1 7sapiens OX = 9606 GN = WARS PE = 1 SV = 2 Vacuolar proteinsorting-associated protein 35 OS = Homo 92 kDa 9 5 0 4 sapiens OX = 9606GN = VPS35 PE = 1 SV = 2 26S proteasome non-ATPase regulatory subunit 361 kDa 8 21 0 6 OS = Homo sapiens OX = 9606 GN = PSMD3 PE = 1 SV = 2 40Sribosomal protein S16 OS = Homo sapiens OX = 9606 16 kDa 8 0 9 7 GN =RPS16 PE = 1 SV = 2 40S ribosomal protein S6 OS = Homo sapiens OX = 960629 kDa 8 0 5 11 GN = RPS6 PE = 1 SV = 1 40S ribosomal protein S9 OS =Homo sapiens OX = 9606 23 kDa 8 54 11 10 GN = RPS9 PE = 1 SV = 3 ADP/ATPtranslocase 1 OS = Homo sapiens OX = 9606 33 kDa 8 0 0 12 GN = SLC25A4PE = 1 SV = 4 Annexin A1 OS = Homo sapiens OX = 9606 GN = ANXA1 39 kDa 80 0 8 PE = 1 SV = 2 ATP-dependent RNA helicase DDX1 OS = Homo sapiens 82kDa 8 15 2 10 OX = 9606 GN = DDX1 PE = 1 SV = 2 Coatomer subunit gamma-1OS = Homo sapiens 98 kDa 8 9 1 6 OX = 9606 GN = COPG1 PE = 1 SV = 1Elongation factor Tu, mitochondrial OS = Homo sapiens 50 kDa 8 0 0 0 OX= 9606 GN = TUFM PE = 1 SV = 2 Erythrocyte band 7 integral membraneprotein OS = Homo 32 kDa 8 6 2 3 sapiens OX = 9606 GN = STOM PE = 1 SV =3 Eukaryotic translation initiation factor 2 subunit 3 51 kDa 8 0 3 12OS = Homo sapiens OX = 9606 GN = EIF2S3 PE = 1 SV = 3 Flotillin-1 OS =Homo sapiens OX = 9606 GN = FLOT1 47 kDa 8 25 1 4 PE = 1 SV = 3Heterogeneous nuclear ribonucleoprotein L OS = Homo 64 kDa 8 11 0 5sapiens OX = 9606 GN = HNRNPL PE = 1 SV = 2 Isoform 2 of Ankycorbin OS =Homo sapiens OX = 9606 110 kDa 8 38 2 5 GN = RAI14 Isoform 2 ofBifunctional purine biosynthesis protein 65 kDa 8 5 0 4 PURH OS = Homosapiens OX = 9606 GN = ATIC Isoform 2 of Eukaryotic translationinitiation factor 5A-1 20 kDa 8 14 1 7 OS = Homo sapiens OX = 9606 GN =EIF5A Isoform 2 of Golgi apparatus protein 1 OS = Homo 137 kDa 8 4 9 1sapiens OX = 9606 GN = GLG1 Isoform 2 of Poly(rC)-binding protein 2 OS =Homo 39 kDa 8 4 0 4 sapiens OX = 9606 GN = PCBP2 Isoform 3 ofHeterogeneous nuclear ribonucleoprotein 33 kDa 8 20 6 11 D0 OS = Homosapiens OX = 9606 GN = HNRNPD Isoform B of Phosphate carrier protein,mitochondrial 40 kDa 8 7 5 8 OS = Homo sapiens OX = 9606 GN = SLC25A3Kinesin-1 heavy chain OS = Homo sapiens OX = 9606 110 kDa 8 5 0 3 GN =KIF5B PE = 1 SV = 1 Malate dehydrogenase, mitochondrial OS = Homosapiens 36 kDa 8 17 0 13 OX = 9606 GN = MDH2 PE = 1 SV = 3 Mitochondrialcarrier homolog 2 OS = Homo sapiens 33 kDa 8 0 2 4 OX = 9606 GN = MTCH2PE = 1 SV = 1 Pre-mRNA-processing factor 19 OS = Homo sapiens 55 kDa 8 00 14 OX = 9606 GN = PRPF19 PE = 1 SV = 1 Prohibitin OS = Homo sapiens OX= 9606 GN = PHB PE = 1 30 kDa 8 14 2 15 SV = 1 Protein transport proteinSec23A OS = Homo sapiens 86 kDa 8 9 1 10 OX = 9606 GN = SEC23A PE = 1 SV= 2 Protein transport protein Sec61 subunit alpha isoform 1 52 kDa 8 2 15 OS = Homo sapiens OX = 9606 GN = SEC61A1 PE = 1 SV = 2 Septin-11 OS =Homo sapiens OX = 9606 GN = SEPT11 49 kDa 8 27 0 9 PE = 1 SV = 3 Smallnuclear ribonucleoprotein Sm D1 OS = Homo 13 kDa 8 0 6 7 sapiens OX =9606 GN = SNRPD1 PE = 1 SV = 1 SWISS-PROT: P15497 (Bos taurus)Apolipoprotein A-I 30 kDa 8 21 6 4 precursor THO complex subunit 4 OS =Homo sapiens OX = 9606 27 kDa 8 37 7 11 GN = ALYREF PE = 1 SV = 3TREMBL: Q3ZBS7 (Bos taurus) Vitronectin 54 kDa 8 26 12 11 Ubiquitin-40Sribosomal protein S27a OS = Homo sapiens 18 kDa 8 0 13 10 OX = 9606 GN =RPS27A PE = 1 SV = 2 26S proteasome non-ATPase regulatory subunit 12 53kDa 7 6 0 2 OS = Homo sapiens OX = 9606 GN = PSMD12 PE = 1 SV = 3 26Sproteasome regulatory subunit 8 OS = Homo sapiens 46 kDa 7 6 0 12 OX =9606 GN = PSMC5 PE = 1 SV = 1 40S ribosomal protein S14 OS = Homosapiens OX = 9606 16 kDa 7 40 13 9 GN = RPS14 PE = 1 SV = 3 40Sribosomal protein S19 OS = Homo sapiens OX = 9606 16 kDa 7 0 12 15 GN =RPS19 PE = 1 SV = 2 40S ribosomal protein S23 OS = Homo sapiens OX =9606 16 kDa 7 0 4 12 GN = RPS23 PE = 1 SV = 3 60S ribosomal protein L30OS = Homo sapiens OX = 9606 13 kDa 7 0 8 8 GN = RPL30 PE = 1 SV = 2ADP-ribosylation factor 3 OS = Homo sapiens OX = 9606 21 kDa 7 11 7 7 GN= ARF3 PE = 1 SV = 2 Aminoacyl tRNA synthase complex-interacting 35 kDa7 0 0 6 multifunctional protein 2 OS = Homo sapiens OX = 9606 GN = AIMP2PE = 1 SV = 2 CAD protein OS = Homo sapiens OX = 9606 GN = CAD 243 kDa 70 0 0 PE = 1 SV = 3 Catenin beta-1 OS = Homo sapiens OX = 9606 85 kDa 73 0 6 GN = CTNNB1 PE = 1 SV = 1 Cell division control protein 42 homologOS = Homo 21 kDa 7 0 2 11 sapiens OX = 9606 GN = CDC42 PE = 1 SV = 2Chloride intracellular channel protein 1 OS = Homo 27 kDa 7 0 0 5sapiens OX = 9606 GN = CLIC1 PE = 1 SV = 4 Coatomer subunit beta OS =Homo sapiens OX = 9606 107 kDa 7 5 0 1 GN = COPB1 PE = 1 SV = 3Eukaryotic initiation factor 4A-III OS = Homo sapiens 47 kDa 7 20 4 8 OX= 9606 GN = EIF4A3 PE = 1 SV = 4 Heterogeneous nuclear ribonucleoproteinH OS = Homo 51 kDa 7 23 1 9 sapiens OX = 9606 GN = HNRNPH1 PE = 1 SV = 1Importin-5 OS = Homo sapiens OX = 9606 GN = IPO5 124 kDa 7 5 0 3 PE = 1SV = 4 Inhibitor of nuclear factor kappa-B kinase-interacting 39 kDa 7 76 7 protein OS = Homo sapiens OX = 9606 GN = IKBIP PE = 1 SV = 1 Isoform2 of 26S proteasome non-ATPase regulatory 48 kDa 7 12 0 5 subunit 11 OS= Homo sapiens OX = 9606 GN = PSMD11 Isoform 2 of F-actin-cappingprotein subunit beta 31 kDa 7 46 10 5 OS = Homo sapiens OX = 9606 GN =CAPZB Isoform 2 of Heterogeneous nuclear ribonucleoprotein D- 34 kDa 7 80 7 like OS = Homo sapiens OX = 9606 GN = HNRNPDL Isoform 2 of Integrinalpha-3 OS = Homo sapiens 119 kDa 7 2 0 5 OX = 9606 GN = ITGA3 Isoform 2of Nucleolar RNA helicase 2 OS = Homo 80 kDa 7 16 2 7 sapiens OX = 9606GN = DDX21 Isoform 2 of PDZ and LIM domain protein 7 OS = Homo 47 kDa 713 0 5 sapiens OX = 9606 GN = PDLIM7 Isoform 2 of Spermine synthase OS =Homo sapiens 35 kDa 7 2 0 6 OX = 9606 GN = SMS Isoform 2 ofTransketolase OS = Homo sapiens OX = 9606 69 kDa 7 29 0 9 GN = TKTIsoform 3 of 116 kDa U5 small nuclear ribonucleoprotein 108 kDa 7 6 0 9component OS = Homo sapiens OX = 9606 GN = EFTUD2 Isoform 3 of 60Sribosomal protein L17 OS = Homo 26 kDa 7 54 5 8 sapiens OX = 9606 GN =RPL17 Isoform 3 of Integrin alpha-V OS = Homo sapiens 111 kDa 7 22 2 12OX = 9606 GN = ITGAV Isoform SV3 of Supervillin OS = Homo sapiens OX =9606 245 kDa 7 46 0 1 GN = SVIL NADH dehydrogenase [ubiquinone] 1 alphasubcomplex 43 kDa 7 12 1 7 subunit 9, mitochondrial OS = Homo sapiens OX= 9606 GN = NDUFA9 PE = 1 SV = 2 Peroxiredoxin-1 OS = Homo sapiens OX =9606 22 kDa 7 0 0 7 GN = PRDX1 PE = 1 SV = 1 Poly(rC)-binding protein 1OS = Homo sapiens OX = 9606 37 kDa 7 0 0 2 GN = PCBP1 PE = 1 SV = 2Pre-mRNA-processing-splicing factor 8 OS = Homo 274 kDa 7 6 1 3 sapiensOX = 9606 GN = PRPF8 PE = 1 SV = 2 Probable ATP-dependent RNA helicaseDDX5 69 kDa 7 25 7 19 OS = Homo sapiens OX = 9606 GN = DDX5 PE = 1 SV =1 Prohibitin-2 OS = Homo sapiens OX = 9606 GN = PHB2 33 kDa 7 25 5 12 PE= 1 SV = 2 Protein disulfide-isomerase A4 OS = Homo sapiens 73 kDa 7 4 04 OX = 9606 GN = PDIA4 PE = 1 SV = 2 Ras GTPase-activatingprotein-binding protein 1 52 kDa 7 19 2 8 OS = Homo sapiens OX = 9606 GN= G3BP1 PE = 1 SV = 1 Ras-related protein Rab-1B OS = Homo sapiens OX =9606 22 kDa 7 0 0 8 GN = RAB1B PE = 1 SV = 1 RNA-binding motif protein,X chromosome OS = Homo 42 kDa 7 22 5 13 sapiens OX = 9606 GN = RBMX PE =1 SV = 3 Splicing factor 3B subunit 1 OS = Homo sapiens 146 kDa 7 16 1 3OX = 9606 GN = SF3B1 PE = 1 SV = 3 SWISS-PROT: P02535-1 Tax_Id = 1009058 kDa 7 39 7 0 Gene_Symbol = Krt10 Isoform 1 of Keratin, type Icytoskeletal 10 SWISS-PROT: P08730-1 Tax_Id = 10090 48 kDa 7 451 17 31Gene_Symbol = Krt13 Isoform 1 of Keratin, type I cytoskeletal 13SWISS-PROT: Q6IFZ6 Tax_Id = 10090 61 kDa 7 32 10 5 Gene_Symbol = Krt77Keratin, type II cytoskeletal 1b Talin-2 OS = Homo sapiens OX = 9606 GN= TLN2 PE = 1 272 kDa 7 0 0 1 SV = 4 Transcription intermediary factor1-beta OS = Homo 89 kDa 7 5 0 0 sapiens OX = 9606 GN = TRIM28 PE = 1 SV= 5 TREMBL: Q9TRI1 (Bos taurus) similar to inter-alpha- 106 kDa 7 57 7 8trypsin inhibitor heavy chain2 U5 small nuclear ribonucleoprotein 200kDa helicase 245 kDa 7 1 0 0 OS = Homo sapiens OX = 9606 GN = SNRNP200PE = 1 SV = 2 Valine--tRNA ligase OS = Homo sapiens OX = 9606 140 kDa 78 0 4 GN = VARS PE = 1 SV = 4 26S proteasome regulatory subunit 4 OS =Homo sapiens 49 kDa 6 25 1 6 OX = 9606 GN = PSMC1 PE = 1 SV = 1 40Sribosomal protein S26 OS = Homo sapiens OX = 9606 13 kDa 6 0 6 6 GN =RPS26 PE = 1 SV = 3 60S ribosomal protein L24 OS = Homo sapiens OX =9606 18 kDa 6 0 5 6 GN = RPL24 PE = 1 SV = 1 60S ribosomal protein L27aOS = Homo sapiens 17 kDa 6 0 6 4 OX = 9606 GN = RPL27A PE = 1 SV = 2Alpha-centractin OS = Homo sapiens OX = 9606 43 kDa 6 26 3 11 GN =ACTR1A PE = 1 SV = 1 ATP-dependent RNA helicase DDX18 OS = Homo 75 kDa 65 3 8 sapiens OX = 9606 GN = DDX18 PE = 1 SV = 2 Collagen alpha-1(IV)chain OS = Homo sapiens OX = 9606 161 kDa 6 69 21 7 GN = COL4A1 PE = 1SV = 4 Collagen alpha-1(VIII) chain OS = Homo sapiens 73 kDa 6 0 29 22OX = 9606 GN = COL8A1 PE = 1 SV = 2 Cullin-associated NEDD8-dissociatedprotein 1 136 kDa 6 0 0 3 OS = Homo sapiens GN = CAND1 PE = 1 SV = 2Dihydrolipoyllysine-residue succinyltransferase 49 kDa 6 17 0 8component of 2-oxoglutarate dehydrogenase complex, mitochondrial OS =Homo sapiens OX = 9606 GN = DLST PE = 1 SV = 4Dihydropyrimidinase-related protein 2 OS = Homo 62 kDa 6 3 0 5 sapiensOX = 9606 GN = DPYSL2 PE = 1 SV = 1 Eukaryotic translation initiationfactor 3 subunit M 43 kDa 6 11 0 5 OS = Homo sapiens OX = 9606 GN =EIF3M PE = 1 SV = 1 Exportin-1 OS = Homo sapiens OX = 9606 GN = XPO1 123kDa 6 0 0 0 PE = 1 SV = 1 F-actin-capping protein subunit alpha-2 OS =Homo 33 kDa 6 33 8 9 sapiens OX = 9606 GN = CAPZA2 PE = 1 SV = 3Glycogen phosphorylase, brain form OS = Homo sapiens 97 kDa 6 0 0 1 OX =9606 GN = PYGB PE = 1 SV = 5 Importin-7 OS = Homo sapiens OX = 9606 GN =IPO7 120 kDa 6 0 0 0 PE = 1 SV = 1 Isoform 2 of 6-phosphogluconatedehydrogenase, 52 kDa 6 4 0 3 decarboxylating OS = Homo sapiens OX =9606 GN = PGD Isoform 2 of Coatomer subunit beta′ OS = Homo sapiens 99kDa 6 10 0 4 OX = 9606 GN = COPB2 Isoform 2 of Coronin-1C OS = Homosapiens OX = 9606 54 kDa 6 36 5 7 GN = CORO1C Isoform 2 of Elongationfactor 1-delta OS = Homo sapiens 71 kDa 6 32 3 4 OX = 9606 GN = EEF1DIsoform 2 of Inverted formin-2 OS = Homo sapiens 135 kDa 6 4 0 1 OX =9606 GN = INF2 Isoform 2 of Programmed cell death 6-interacting protein97 kDa 6 3 0 4 OS = Homo sapiens OX = 9606 GN = PDCD6IP Isoform 2 ofSurfeit locus protein 4 OS = Homo sapiens 18 kDa 6 10 3 7 OX = 9606 GN =SURF4 Isoform 3 of Plasminogen activator inhibitor 1 RNA- 43 kDa 6 11 67 binding protein OS = Homo sapiens OX = 9606 GN = SERBP1 Isoform 5 ofPhosphatidylinositol-binding clathrin 70 kDa 6 5 0 7 assembly protein OS= Homo sapiens GN = PICALM Junction plakoglobin OS = Homo sapiens OX =9606 82 kDa 6 38 0 6 GN = JUP PE = 1 SV = 3 Lamina-associatedpolypeptide 2, isoforms beta/gamma 51 kDa 6 26 7 7 OS = Homo sapiens OX= 9606 GN = TMPO PE = 1 SV = 2 Leucine-rich repeat-containing protein 59OS = Homo 35 kDa 6 0 7 10 sapiens OX = 9606 GN = LRRC59 PE = 1 SV = 1Multifunctional protein ADE2 OS = Homo sapiens 47 kDa 6 3 0 2 OX = 9606GN = PAICS PE = 1 SV = 3 Nicotinamide N-methyltransferase OS = Homosapiens 30 kDa 6 0 0 1 OX = 9606 GN = NNMT PE = 1 SV = 1 Poly[ADP-ribose] polymerase 4 OS = Homo sapiens 193 kDa 6 19 0 5 OX = 9606GN = PARP4 PE = 1 SV = 3 Protein S100-A6 OS = Homo sapiens OX = 9606 10kDa 6 0 1 4 GN = S100A6 PE = 1 SV = 1 RuvB-like 2 OS = Homo sapiens OX =9606 GN = RUVBL2 51 kDa 6 19 0 5 PE = 1 SV = 3 Sarcoplasmic/endoplasmicreticulum calcium ATPase 2 115 kDa 6 6 0 0 OS = Homo sapiens GN = ATP2A2PE = 1 SV = 1 Signal transducer and activator of transcription 1- 87 kDa6 15 1 5 alpha/beta OS = Homo sapiens OX = 9606 GN = STAT1 PE = 1 SV = 2Synaptic vesicle membrane protein VAT-1 homolog 42 kDa 6 6 0 7 OS = Homosapiens OX = 9606 GN = VAT1 PE = 1 SV = 2 Ubiquitin carboxyl-terminalhydrolase OS = Homo sapiens 27 kDa 6 0 0 4 OX = 9606 GN = UCHL1 PE = 1SV = 1 Very-long-chain 3-oxoacyl-CoA reductase OS = Homo 34 kDa 6 0 2 6sapiens OX = 9606 GN = HSD17B12 PE = 1 SV = 2 26S proteasome non-ATPaseregulatory subunit 13 43 kDa 5 9 0 4 OS = Homo sapiens OX = 9606 GN =PSMD13 PE = 1 SV = 2 26S proteasome regulatory subunit 10B OS = Homo 44kDa 5 0 0 5 sapiens OX = 9606 GN = PSMC6 PE = 1 SV = 1 40S ribosomalprotein S10 OS = Homo sapiens OX = 9606 19 kDa 5 38 13 12 GN = RPS10 PE= 1 SV = 1 40S ribosomal protein S11 OS = Homo sapiens OX = 9606 18 kDa5 26 3 7 GN = RPS11 PE = 1 SV = 3 60S ribosomal protein L21 OS = Homosapiens OX = 9606 19 kDa 5 34 8 7 GN = RPL21 PE = 1 SV = 2 60S ribosomalprotein L22 OS = Homo sapiens OX = 9606 15 kDa 5 0 6 8 GN = RPL22 PE = 1SV = 2 60S ribosomal protein L23a (Fragment) OS = Homo 19 kDa 5 0 9 9sapiens OX = 9606 GN = RPL23A PE = 1 SV = 1 60S ribosomal protein L27 OS= Homo sapiens OX = 9606 16 kDa 5 0 6 11 GN = RPL27 PE = 1 SV = 2 60Sribosomal protein L28 OS = Homo sapiens OX = 9606 16 kDa 5 13 4 4 GN =RPL28 PE = 1 SV = 3 60S ribosomal protein L31 OS = Homo sapiens OX =9606 14 kDa 5 33 8 6 GN = RPL31 PE = 1 SV = 1 60S ribosomal protein L36OS = Homo sapiens OX = 9606 12 kDa 5 0 7 7 GN = RPL36 PE = 1 SV = 3Actin-related protein 2/3 complex subunit 3 OS = Homo 21 kDa 5 0 4 7sapiens OX = 9606 GN = ARPC3 PE = 1 SV = 3 Actin-related protein 2/3complex subunit 4 OS = Homo 20 kDa 5 19 2 6 sapiens OX = 9606 GN = ARPC4PE = 1 SV = 3 Calpain-1 catalytic subunit OS = Homo sapiens OX = 9606 82kDa 5 27 0 6 GN = CAPN1 PE = 1 SV = 1 Calponin-2 OS = Homo sapiens OX =9606 GN = CNN2 34 kDa 5 5 1 6 PE = 1 SV = 4 Cathepsin D OS = Homosapiens OX = 9606 GN = CTSD 45 kDa 5 0 0 3 PE = 1 SV = 1 Cleavage andpolyadenylation specificity factor subunit 5 26 kDa 5 0 0 6 OS = Homosapiens OX = 9606 GN = NUDT21 PE = 1 SV = 1 Coatomer subunit epsilon OS= Homo sapiens GN = COPE 34 kDa 5 14 2 8 PE = 1 SV = 3 Copine-3 OS =Homo sapiens OX = 9606 GN = CPNE3 60 kDa 5 0 0 1 PE = 1 SV = 1Cytochrome c oxidase subunit 2 OS = Homo sapiens 26 kDa 5 0 0 6 OX =9606 GN = MT-CO2 PE = 1 SV = 1 DNA-(apurinic or apyrimidinic site) lyaseOS = Homo 36 kDa 5 8 0 3 sapiens OX = 9606 GN = APEX1 PE = 1 SV = 2Erlin-1 OS = Homo sapiens OX = 9606 GN = ERLIN1 PE = 1 39 kDa 5 0 0 4 SV= 1 Eukaryotic translation elongation factor 1 epsilon-1 20 kDa 5 7 2 7OS = Homo sapiens GN = EEF1E1 PE = 1 SV = 1 Eukaryotic translationinitiation factor 2 subunit 1 36 kDa 5 9 1 7 OS = Homo sapiens OX = 9606GN = EIF2S1 PE = 1 SV = 3 Eukaryotic translation initiation factor 3subunit H 42 kDa 5 0 0 3 OS = Homo sapiens OX = 9606 GN = EIF3H PE = 1SV = 1 Heat shock 70 kDa protein 4 OS = Homo sapiens 94 kDa 5 4 0 1 OX =9606 GN = HSPA4 PE = 1 SV = 4 High mobility group protein B1 OS = Homosapiens 25 kDa 5 0 0 3 OX = 9606 GN = HMGB1 PE = 1 SV = 3 Importin-9 OS= Homo sapiens OX = 9606 GN = IPO9 116 kDa 5 0 0 4 PE = 1 SV = 3 Isoform2 of 26S proteasome non-ATPase regulatory 102 kDa 5 10 0 3 subunit 1 OS= Homo sapiens OX = 9606 GN = PSMD1 Isoform 2 of Calcium-bindingmitochondrial carrier 74 kDa 5 10 0 9 protein Aralar2 OS = Homo sapiensOX = 9606 GN = SLC25A13 Isoform 2 of Collagen alpha-3(VI) chain OS =Homo 321 kDa 5 0 1 1 sapiens OX = 9606 GN = COL6A3 Isoform 2 ofEukaryotic translation initiation factor 3 99 kDa 5 5 0 6 subunit B OS =Homo sapiens OX = 9606 GN = EIF3B Isoform 2 of Glucose-6-phosphateisomerase OS = Homo 64 kDa 5 4 0 2 sapiens OX = 9606 GN = GPI Isoform 2of HLA class I histocompatibility antigen, A- 41 kDa 5 3 0 3 11 alphachain OS = Homo sapiens OX = 9606 GN = HLA- A Isoform 2 of Myb-bindingprotein 1A OS = Homo sapiens 149 kDa 5 1 0 0 OX = 9606 GN = MYBBP1AIsoform 2 of Serine/arginine-rich splicing factor 2 24 kDa 5 2 0 2 OS =Homo sapiens OX = 9606 GN = SRSF2 Isoform 2 of U1 small nuclearribonucleoprotein 70 kDa 51 kDa 5 0 0 5 OS = Homo sapiens OX = 9606 GN =SNRNP70 Isoform 3 of Glutaminase kidney isoform, mitochondrial 65 kDa 57 0 3 OS = Homo sapiens OX = 9606 GN = GLS Isoform 4 of AP-3 complexsubunit delta-1 OS = Homo 115 kDa 5 5 0 2 sapiens GN = AP3D1 Isoform 4of Protein phosphatase 1 regulatory subunit 109 kDa 5 10 3 7 12A OS =Homo sapiens OX = 9606 GN = PPP1R12A Isoform Short of Eukaryotictranslation initiation factor 25 kDa 5 5 1 4 4H OS = Homo sapiens OX =9606 GN = EIF4H Lon protease homolog, mitochondrial OS = Homo sapiens106 kDa 5 0 0 6 GN = LONP1 PE = 1 SV = 2 Mannosyl-oligosaccharideglucosidase OS = Homo 92 kDa 5 22 4 11 sapiens GN = MOGS PE = 1 SV = 5Prolyl 3-hydroxylase 1 OS = Homo sapiens OX = 9606 83 kDa 5 6 0 3 GN =P3H1 PE = 1 SV = 2 Proteasome subunit alpha type-4 OS = Homo sapiens 29kDa 5 0 0 7 OX = 9606 GN = PSMA4 PE = 1 SV = 1 Protein LYRIC OS = Homosapiens OX = 9606 64 kDa 5 10 6 9 GN = MTDH PE = 1 SV = 2Protein/nucleic acid deglycase DJ-1 OS = Homo sapiens 20 kDa 5 0 0 3 OX= 9606 GN = PARK7 PE = 1 SV = 2 Ras-related protein Rab-1A OS = Homosapiens OX = 9606 23 kDa 5 13 2 8 GN = RAB1A PE = 1 SV = 3 Ras-relatedprotein Rab-3B OS = Homo sapiens OX = 9606 25 kDa 5 0 2 7 GN = RAB3B PE= 1 SV = 2 Rho GDP-dissociation inhibitor 1 OS = Homo sapiens 23 kDa 5 00 4 OX = 9606 GN = ARHGDIA PE = 1 SV = 3 Signal recognition particlesubunit SRP72 OS = Homo 75 kDa 5 8 1 8 sapiens OX = 9606 GN = SRP72 PE =1 SV = 3 Small nuclear ribonucleoprotein Sm D2 OS = Homo 14 kDa 5 11 0 5sapiens OX = 9606 GN = SNRPD2 PE = 1 SV = 1 Sorting and assemblymachinery component 50 homolog 52 kDa 5 19 4 12 OS = Homo sapiens OX =9606 GN = SAMM50 PE = 1 SV = 3 Src substrate cortactin OS = Homo sapiensOX = 9606 62 kDa 5 38 12 8 GN = CTTN PE = 1 SV = 2 SWI/SNF complexsubunit SMARCC1 OS = Homo 123 kDa 5 0 0 2 sapiens OX = 9606 GN = SMARCC1PE = 1 SV = 3 Transcription factor BTF3 OS = Homo sapiens OX = 9606 22kDa 5 8 0 7 GN = BTF3 PE = 1 SV = 1 Translocon-associated proteinsubunit delta OS = Homo 19 kDa 5 0 2 4 sapiens OX = 9606 GN = SSR4 PE =1 SV = 1 TREMBL: Q1RMK2 (Bos taurus) IGHM protein 65 kDa 5 0 0 0 WDrepeat-containing protein 1 OS = Homo sapiens 66 kDa 5 11 0 5 OX = 9606GN = WDR1 PE = 1 SV = 4 Zyxin OS = Homo sapiens OX = 9606 GN = ZYX PE =1 61 kDa 5 13 0 4 SV = 1 2-oxoglutarate dehydrogenase, mitochondrial OS= Homo 116 kDa 4 2 0 6 sapiens OX = 9606 GN = OGDH PE = 1 SV = 3 40Sribosomal protein S27-like OS = Homo sapiens 9 kDa 4 0 1 1 OX = 9606 GN= RPS27L PE = 1 SV = 3 Adenosylhomocysteinase OS = Homo sapiens OX =9606 48 kDa 4 4 0 2 GN = AHCY PE = 1 SV = 4 ADP-ribosylation factor 6 OS= Homo sapiens OX = 9606 20 kDa 4 0 0 2 GN = ARF6 PE = 1 SV = 2Alanine--tRNA ligase, cytoplasmic OS = Homo sapiens 107 kDa 4 0 0 2 OX =9606 GN = AARS PE = 1 SV = 2 ATPase family AAA domain-containing protein3A 71 kDa 4 22 5 9 OS = Homo sapiens OX = 9606 GN = ATAD3A PE = 1 SV = 2Coatomer subunit delta OS = Homo sapiens OX = 9606 57 kDa 4 10 0 5 GN =ARCN1 PE = 1 SV = 1 Cold-inducible RNA-binding protein OS = Homo sapiens19 kDa 4 3 0 4 GN = CIRBP PE = 1 SV = 1 Copine-1 OS = Homo sapiens OX =9606 GN = CPNE1 59 kDa 4 0 0 2 PE = 1 SV = 1 Cytochrome b-c1 complexsubunit 2, mitochondrial 48 kDa 4 0 0 8 OS = Homo sapiens OX = 9606 GN =UQCRC2 PE = 1 SV = 3 DBH-like monooxygenase protein 1 OS = Homo sapiens70 kDa 4 17 5 4 OX = 9606 GN = MOXD1 PE = 1 SV = 1 EH domain-containingprotein 1 OS = Homo sapiens 61 kDa 4 11 0 9 OX = 9606 GN = EHD1 PE = 1SV = 2 Eukaryotic translation initiation factor 3 subunit C 105 kDa 4 40 5 OS = Homo sapiens OX = 9606 GN = EIF3C PE = 1 SV = 1 Eukaryotictranslation initiation factor 4 gamma 2 102 kDa 4 8 1 3 OS = Homosapiens GN = EIF4G2 PE = 1 SV = 1 Eukaryotic translation initiationfactor 6 OS = Homo 27 kDa 4 10 2 5 sapiens OX = 9606 GN = EIF6 PE = 1 SV= 1 Guanine nucleotide-binding protein G(I)/G(S)/G(T) 37 kDa 4 12 3 4subunit beta-1 OS = Homo sapiens OX = 9606 GN = GNB1 PE = 1 SV = 3Isoform 10 of Calpastatin OS = Homo sapiens OX = 9606 82 kDa 4 4 0 1 GN= CAST Isoform 2 of 40S ribosomal protein S24 OS = Homo 15 kDa 4 46 10 8sapiens OX = 9606 GN = RPS24 Isoform 2 ofChromodomain-helicase-DNA-binding 221 kDa 4 5 0 1 protein 4 OS = Homosapiens GN = CHD4 Isoform 2 of Eukaryotic translation initiation factor3 61 kDa 4 5 0 6 subunit L OS = Homo sapiens OX = 9606 GN = EIF3LIsoform 2 of Glucosidase 2 subunit beta OS = Homo 59 kDa 4 5 0 4 sapiensOX = 9606 GN = PRKCSH Isoform 2 of Hexokinase-1 OS = Homo sapiens OX =9606 102 kDa 4 3 0 1 GN = HK1 Isoform 2 of Isocitrate dehydrogenase[NADP], 45 kDa 4 19 1 4 mitochondrial OS = Homo sapiens OX = 9606 GN =IDH2 Isoform 2 of Nodal modulator 2 OS = Homo sapiens 134 kDa 4 4 0 3 OX= 9606 GN = NOMO2 Isoform 2 of Plastin-3 OS = Homo sapiens OX = 9606 69kDa 4 0 0 1 GN = PLS3 Isoform 2 of Proteasome subunit alpha type-3 OS =Homo 28 kDa 4 6 0 4 sapiens OX = 9606 GN = PSMA3 Isoform 2 of ProteinSET OS = Homo sapiens GN = SET 32 kDa 4 7 0 4 Isoform 2 of Splicingfactor U2AF 65 kDa subunit 53 kDa 4 1 0 5 OS = Homo sapiens OX = 9606 GN= U2AF2 Isoform 2 of SWI/SNF complex subunit SMARCC2 125 kDa 4 9 0 3 OS= Homo sapiens OX = 9606 GN = SMARCC2 Isoform 2 of Unconventionalmyosin-Ib OS = Homo 125 kDa 4 16 0 0 sapiens OX = 9606 GN = MYO1BIsoform 3 of Dynactin subunit 1 OS = Homo sapiens 137 kDa 4 8 0 2 OX =9606 GN = DCTN1 Isoform 3 of Heterogeneous nuclear ribonucleoprotein 31kDa 4 10 2 6 A/B OS = Homo sapiens GN = HNRNPAB Isoform 3 of MyoferlinOS = Homo sapiens OX = 9606 233 kDa 4 6 0 1 GN = MYOF Isoform 3 ofNucleolar and coiled-body phosphoprotein 1 74 kDa 4 7 1 3 OS = Homosapiens OX = 9606 GN = NOLC1 Isoform 4 of Inhibitor of nuclear factorkappa-B kinase- 43 kDa 4 11 2 5 interacting protein OS = Homo sapiens OX= 9606 GN = IKBIP Isoform 6 of MMS19 nucleotide excision repair protein108 kDa 4 0 0 1 homolog OS = Homo sapiens GN = MMS19 Isoform Short ofRNA-binding protein FUS OS = Homo 53 kDa 4 9 0 4 sapiens OX = 9606 GN =FUS Microtubule-actin cross-linking factor 1, isoforms 1/2/3/5 838 kDa 42 0 0 OS = Homo sapiens OX = 9606 GN = MACF1 PE = 1 SV = 4 Mitochondrialcarrier homolog 1 (Fragment) OS = Homo 43 kDa 4 0 0 5 sapiens OX = 9606GN = MTCH1 PE = 1 SV = 1 Nuclease-sensitive element-binding protein 1 OS= Homo 36 kDa 4 0 7 9 sapiens OX = 9606 GN = YBX1 PE = 1 SV = 3Nucleolar GTP-binding protein 1 OS = Homo sapiens 74 kDa 4 5 0 1 OX =9606 GN = GTPBP4 PE = 1 SV = 3 OCIA domain-containing protein 2 OS =Homo sapiens 17 kDa 4 0 0 0 OX = 9606 GN = OCIAD2 PE = 1 SV = 1Peroxisomal multifunctional enzyme type 2 OS = Homo 80 kDa 4 10 0 9sapiens GN = HSD17B4 PE = 1 SV = 3 Probable ATP-dependent RNA helicaseDDX6 54 kDa 4 4 0 2 OS = Homo sapiens OX = 9606 GN = DDX6 PE = 1 SV = 2Proteasome subunit alpha type-2 OS = Homo sapiens 26 kDa 4 0 0 8 OX =9606 GN = PSMA2 PE = 1 SV = 2 Proteasome subunit beta type-1 OS = Homosapiens 26 kDa 4 0 0 6 OX = 9606 GN = PSMB1 PE = 1 SV = 2 Proteasomesubunit beta type-3 OS = Homo sapiens 23 kDa 4 0 0 6 OX = 9606 GN =PSMB3 PE = 1 SV = 2 Proteasome subunit beta type-7 OS = Homo sapiens 30kDa 4 0 0 6 OX = 9606 GN = PSMB7 PE = 1 SV = 1 Protein arginineN-methyltransferase 1 OS = Homo 42 kDa 4 3 0 1 sapiens GN = PRMT1 PE = 1SV = 2 Ras GTPase-activating protein-binding protein 2 54 kDa 4 14 2 5OS = Homo sapiens OX = 9606 GN = G3BP2 PE = 1 SV = 2 Ras suppressorprotein 1 OS = Homo sapiens OX = 9606 32 kDa 4 16 1 8 GN = RSU1 PE = 1SV = 3 RNA-binding protein Raly OS = Homo sapiens OX = 9606 32 kDa 4 142 7 GN = RALY PE = 1 SV = 1 Sideroflexin-1 OS = Homo sapiens OX = 9606GN= SFXN1 36 kDa 4 3 1 4 PE = 1 SV = 4 Sideroflexin-3 OS = Homo sapiens OX= 9606GN = SFXN3 36 kDa 4 0 0 4 PE = 1 SV = 3 Signal recognitionparticle 9 kDa protein OS = Homo 10 kDa 4 7 1 3 sapiens OX = 9606 GN =SRP9 PE = 1 SV = 2 Splicing factor 3A subunit 1 OS = Homo sapiens 89 kDa4 6 0 5 OX = 9606 GN = SF3A1 PE = 1 SV = 1 SWISS-PROT: Q9TTE1 (Bostaurus) Endopin-1 46 kDa 4 4 2 1 precursor Thy-l membrane glycoproteinOS = Homo sapiens 18 kDa 4 0 6 3 OX = 9606 GN = THY1 PE = 1 SV = 2Transmembrane protein 43 OS = Homo sapiens OX = 9606 45 kDa 4 0 5 6 GN =TMEM43 PE = 1 SV = 1 TREMBL: Q1A7A4 (Bos taurus) similar to complement189 kDa 4 0 0 0 component C5 Tricarboxylate transport protein,mitochondrial 34 kDa 4 2 0 3 OS = Homo sapiens OX = 9606 GN = SLC25A1 PE= 1 SV = 2 U2 small nuclear ribonucleoprotein A′ OS = Homo sapiens 28kDa 4 0 0 4 OX = 9606 GN = SNRPA1 PE = 1 SV = 2 Vasodilator-stimulatedphosphoprotein OS = Homo 40 kDa 4 10 0 10 sapiens OX = 9606 GN = VASP PE= 1 SV = 3 (Bos taurus) 47 kDa protein 47 kDa 3 0 0 0 (Bos taurus)similar to Complement C4-A precursor 193 kDa 3 7 4 4 26S proteasomenon-ATPase regulatory subunit 5 56 kDa 3 4 0 1 OS = Homo sapiens OX =9606 GN = PSMD5 PE = 1 SV = 3 26S proteasome non-ATPase regulatorysubunit 7 37 kDa 3 10 0 6 OS = Homo sapiens OX = 9606 GN = PSMD7 PE = 1SV = 2 26S proteasome regulatory subunit 7 OS = Homo sapiens 49 kDa 3 40 4 OX = 9606 GN = PSMC2 PE = 1 SV = 3 39S ribosomal protein L28,mitochondrial OS = Homo 30 kDa 3 0 0 1 sapiens OX = 9606 GN = MRPL28 PE= 1 SV = 4 40S ribosomal protein S15a OS = Homo sapiens OX = 9606 15 kDa3 0 2 3 GN = RPS15A PE = 1 SV = 2 60S ribosomal protein L15 OS = Homosapiens OX = 9606 24 kDa 3 27 3 11 GN = RPL15 PE = 1 SV = 2 Acetyl-CoAacetyltransferase, mitochondrial OS = Homo 45 kDa 3 11 0 5 sapiens OX =9606 GN = ACAT1 PE = 1 SV = 1 ATP synthase F(0) complex subunit B1,mitochondrial 29 kDa 3 0 0 2 OS = Homo sapiens OX = 9606 GN = ATP5PB PE= 1 SV = 2 ATP-binding cassette sub-family D member 3 OS = Homo 75 kDa 38 2 2 sapiens OX = 9606 GN = ABCD3 PE = 1 SV = 1 Calponin-3 OS = Homosapiens OX = 9606 GN = CNN3 36 kDa 3 4 1 6 PE = 1 SV = 1 Cell surfaceglycoprotein MUC18 OS = Homo sapiens 72 kDa 3 3 0 6 OX = 9606 GN = MCAMPE = 1 SV = 2 CTP synthase 1 OS = Homo sapiens OX = 9606 67 kDa 3 0 0 0GN = CTPS1 PE = 1 SV = 2 Cytoplasmic dynein 1 light intermediate chain 157 kDa 3 11 0 4 OS = Homo sapiens OX = 9606 GN = DYNC1LI1 PE = 1 SV = 3Cytoplasmic FMR1-interacting protein 1 OS = Homo 145 kDa 3 2 0 0 sapiensOX = 9606 GN = CYFIP1 PE = 1 SV = 1 Desmoglein-2 OS = Homo sapiens OX =9606 GN = DSG2 122 kDa 3 17 5 7 PE = 1 SV = 2 Desmoplakin OS = Homosapiens OX = 9606 GN = DSP 332 kDa 3 35 0 0 PE = 1 SV = 3Dihydrolipoyllysine-residue acetyltransferase component 69 kDa 3 0 2 10of pyruvate dehydrogenase complex, mitochondrial OS = Homo sapiens OX =9606 GN = DLAT PE = 1 SV = 3 DnaJ homolog subfamily A member 2 OS = Homosapiens 46 kDa 3 0 1 6 OX = 9606 GN = DNAJA2 PE = 1 SV = 1 Dynein lightchain Tctex-type 1 OS = Homo sapiens 12 kDa 3 13 3 5 OX = 9606 GN =DYNLT1 PE = 1 SV = 1 E3 ubiquitin/ISG15 ligase TRIM25 OS = Homo sapiens71 kDa 3 0 0 1 OX = 9606 GN = TRIM25 PE = 1 SV = 2 EGF-like repeat anddiscoidin I-like domain-containing 54 kDa 3 26 1 7 protein 3 OS = Homosapiens OX = 9606 GN = EDIL3 PE = 1 SV = 1 Enoyl-CoA hydratase,mitochondrial OS = Homo sapiens 31 kDa 3 0 0 2 OX = 9606 GN = ECHS1 PE =1 SV = 4 Epidermal growth factor receptor kinase substrate 8-like 81 kDa3 4 0 5 protein 2 OS = Homo sapiens OX = 9606 GN = EPS8L2 PE = 1 SV = 2ER lumen protein-retaining receptor 1 OS = Homo sapiens 25 kDa 3 0 2 1OX = 9606 GN = KDELR1 PE = 1 SV = 1 Eukaryotic translation initiationfactor 2 subunit 2 38 kDa 3 0 7 10 OS = Homo sapiens OX = 9606 GN =EIF2S2 PE = 1 SV = 2 General transcription factor II-I OS = Homo sapiens112 kDa 3 27 3 3 OX = 9606 GN = GTF2I PE = 1 SV = 2 High mobility groupprotein HMGI-C OS = Homo sapiens 12 kDa 3 48 3 0 GN = HMGA2 PE = 1 SV =1 Inosine-5′-monophosphate dehydrogenase 2 OS = Homo 56 kDa 3 0 0 2sapiens OX = 9606 GN = IMPDH2 PE = 1 SV = 2 Isoform 1 of Apoptosisinhibitor 5 OS = Homo sapiens 49 kDa 3 5 0 3 OX = 9606 GN = API5 Isoform12 of Titin OS = Homo sapiens OX = 9606 3994 kDa 3 1 3 3 GN = TTNIsoform 2 of 3-hydroxyacyl-CoA dehydrogenase type-2 26 kDa 3 2 0 1 OS =Homo sapiens OX = 9606 GN = HSD17B10 Isoform 2 of ATP-dependent RNAhelicase DDX54 99 kDa 3 1 0 3 OS = Homo sapiens OX = 9606 GN = DDX54Isoform 2 of B-cell receptor-associated protein 31 35 kDa 3 1 4 7 OS =Homo sapiens OX = 9606 GN = BCAP31 Isoform 2 of Calcium-bindingmitochondrial carrier 51 kDa 3 1 0 5 protein SCaMC-1 OS = Homo sapiensOX = 9606 GN = SLC25A24 Isoform 2 of Collagen alpha-1(V) chain OS = Homo184 kDa 3 0 13 6 sapiens OX = 9606 GN = COL5A1 Isoform 2 of E3ubiquitin-protein ligase UBR4 576 kDa 3 0 0 1 OS = Homo sapiens OX =9606 GN = UBR4 Isoform 2 of Importin-4 OS = Homo sapiens GN = IPO4 119kDa 3 0 0 0 Isoform 2 of Insulin-like growth factor 2 mRNA-binding 62kDa 3 19 2 5 protein 2 OS = Homo sapiens GN = IGF2BP2 Isoform 2 of NADHdehydrogenase [ubiquinone] 1 alpha 49 kDa 3 0 0 2 subcomplex subunit 10,mitochondrial OS = Homo sapiens OX = 9606 GN = NDUFA10 Isoform 2 ofNADH-cytochrome b5 reductase 3 32 kDa 3 6 0 3 OS = Homo sapiens OX =9606 GN = CYB5R3 Isoform 2 of Nuclear pore complex protein Nup107 103kDa 3 1 0 0 OS = Homo sapiens OX = 9606 GN = NUP107 Isoform 2 of ProteinDok-7 OS = Homo sapiens OX = 9606 37 kDa 3 0 2 5 GN = DOK7 Isoform 2 ofProtein FAM98B OS = Homo sapiens 46 kDa 3 9 0 4 OX = 9606 GN = FAM98BIsoform 2 of Sacsin OS = Homo sapiens OX = 9606 437 kDa 3 0 0 0 GN =SACS Isoform 2 of Serine/arginine-rich splicing factor 3 14 kDa 3 8 0 4OS = Homo sapiens OX = 9606 GN = SRSF3 Isoform 2 of Small nuclearribonucleoprotein Sm D3 13 kDa 3 10 2 3 OS = Homo sapiens OX = 9606 GN =SNRPD3 Isoform 2 of Stathmin OS = Homo sapiens OX = 9606 20 kDa 3 0 0 3GN = STMN1 Isoform 2 of TP53-binding protein 1 OS = Homo sapiens 214 kDa3 4 0 0 OX = 9606 GN = TP53BP1 Isoform 2 of Very long-chain specificacyl-CoA 68 kDa 3 10 0 5 dehydrogenase, mitochondrial OS = Homo sapiensOX = 9606 GN = ACADVL Isoform 2 of Voltage-dependent anion-selectivechannel 31 kDa 3 15 0 5 protein 3 OS = Homo sapiens OX = 9606 GN = VDAC3Isoform 2 of V-type proton ATPase catalytic subunit A 65 kDa 3 15 1 2 OS= Homo sapiens OX = 9606 GN = ATP6V1A Isoform 3 of 4F2 cell-surfaceantigen heavy chain 62 kDa 3 1 1 1 OS = Homo sapiens GN = SLC3A2 Isoform3 of Aldehyde dehydrogenase family 16 member 80 kDa 3 0 0 1 A1 OS = Homosapiens OX = 9606 GN = ALDH16A1 Isoform 3 of Drebrin OS = Homo sapiensGN = DBN1 76 kDa 3 31 4 3 Isoform 3 of Erbin OS = Homo sapiens OX = 9606153 kDa 3 0 0 0 GN = ERBIN Isoform 3 of Nucleoside diphosphate kinase BOS = Homo 30 kDa 3 12 0 7 sapiens GN = NME2 Isoform 3 of Perilipin-3 OS= Homo sapiens OX = 9606 47 kDa 3 1 0 3 GN = PLIN3 Isoform 3 of SUNdomain-containing protein 2 80 kDa 3 15 4 12 OS = Homo sapiens OX = 9606GN = SUN2 Isoform 3 of Transportin-3 OS = Homo sapiens OX = 9606 103 kDa3 0 0 0 GN = TNPO3 Isoform 4 of 26S proteasome non-ATPase regulatory 52kDa 3 17 0 2 subunit 6 OS = Homo sapiens OX = 9606 GN = PSMD6 Isoform 9of Protein transport protein Sec31A OS = Homo 131 kDa 3 3 0 1 sapiens GN= SEC31A Isoform K of Kinesin light chain 1 OS = Homo sapiens 70 kDa 3 20 2 OX = 9606 GN = KLC1 MICOS complex subunit MIC19 OS = Homo sapiens 26kDa 3 0 3 6 OX = 9606 GN = CHCHD3 PE = 1 SV = 1 Nuclear pore complexprotein Nup205 OS = Homo 228 kDa 3 0 0 0 sapiens OX = 9606 GN = NUP205PE = 1 SV = 3 Nuclear pore complex protein Nup93 OS = Homo sapiens 93kDa 3 6 0 3 OX = 9606 GN = NUP93 PE = 1 SV = 2 PC4 and SFRS1-interacting protein OS = Homo sapiens 60 kDa 3 16 1 5 OX = 9606 GN =PSIP1 PE = 1 SV = 1 Peroxiredoxin-6 OS = Homo sapiens OX = 9606 25 kDa 30 0 0 GN = PRDX6 PE = 1 SV = 3 PRA1 family protein 3 OS = Homo sapiensOX = 9606 22 kDa 3 7 1 4 GN = ARL6IP5 PE = 1 SV = 1 Proliferating cellnuclear antigen OS = Homo sapiens 29 kDa 3 0 0 0 OX = 9606 GN = PCNA PE= 1 SV = 1 Proliferation-associated protein 2G4 OS = Homo sapiens 44 kDa3 16 0 6 OX = 9606 GN = PA2G4 PE = 1 SV = 3 Proteasome adapter andscaffold protein ECM29 204 kDa 3 0 0 2 OS = Homo sapiens OX = 9606 GN =ECPAS PE = 1 SV = 2 Proteasome subunit beta type-4 OS = Homo sapiens 29kDa 3 0 0 4 OX = 9606 GN = PSMB4 PE = 1 SV = 4 Proteasome subunit betatype-6 OS = Homo sapiens 25 kDa 3 0 0 3 OX = 9606 GN = PSMB6 PE = 1 SV =4 Protein DEK OS = Homo sapiens OX = 9606 GN = DEK 43 kDa 3 2 0 2 PE = 1SV = 1 Protein S 100-A11 OS = Homo sapiens OX = 9606 12 kDa 3 0 0 2 GN =S100A11 PE = 1 SV = 2 Protein transport protein Sec61 subunit beta OS =Homo 10 kDa 3 1 2 2 sapiens OX = 9606 GN = SEC61B PE = 1 SV = 2Puromycin-sensitive aminopeptidase OS = Homo sapiens 103 kDa 3 0 0 2 GN= NPEPPS PE = 1 SV = 2 Ras-related protein Rab-11B OS = Homo sapiens 24kDa 3 9 1 6 OX = 9606 GN = RAB11B PE = 1 SV = 4 Ras-related proteinRab-14 OS = Homo sapiens OX = 9606 24 kDa 3 0 0 6 GN = RAB14 PE = 1 SV =4 Ras-related protein Rap-1b OS = Homo sapiens OX = 9606 21 kDa 3 7 2 3GN = RAP1B PE = 1 SV = 1 Ras-related protein R-Ras OS = Homo sapiens OX= 9606 23 kDa 3 0 0 2 GN = RRAS PE = 1 SV = 1 RNA transcription,translation and transport factor 28 kDa 3 0 4 6 protein OS = Homosapiens OX = 9606 GN = RTRAF PE = 1 SV = 1 SAP domain-containingribonucleoprotein OS = Homo 24 kDa 3 0 0 2 sapiens OX = 9606 GN = SARNPPE = 1 SV = 3 SWISS-PROT: P06868 (Bos taurus) Plasminogen 91 kDa 3 1 5 0precursor SWISS-PROT: P41361 (Bos taurus) Antithrombin-III 52 kDa 3 3 00 precursor TAR DNA-binding protein 43 OS = Homo sapiens 45 kDa 3 2 0 3OX = 9606 GN = TARDBP PE = 1 SV = 1 Thioredoxin reductase 1, cytoplasmicOS = Homo sapiens 71 kDa 3 0 0 1 GN = TXNRD1 PE = 1 SV = 3Thrombospondin type-1 domain-containing protein 4 112 kDa 3 22 10 1 OS =Homo sapiens OX = 9606 GN = THSD4 PE = 2 SV = 2 TREMBL: Q2KJF1 (Bostaurus) Alpha-1-Bglycoprotein 54 kDa 3 11 1 3 tRNA-splicing ligase RtcBhomolog OS = Homo sapiens 55 kDa 3 19 3 9 OX = 9606 GN = RTCB PE = 1 SV= 1 Tyrosine--tRNA ligase, cytoplasmic OS = Homo sapiens 59 kDa 3 0 0 2OX = 9606 GN = YARS PE = 1 SV = 4 Ubiquitin thioesterase OTUB1 OS = Homosapiens 31 kDa 3 0 0 1 OX = 9606 GN = OTUB1 PE = 1 SV = 2 Vigilin OS =Homo sapiens OX = 9606 GN = HDLBP PE = 1 141 kDa 3 9 0 2 SV = 2 V-typeproton ATPase 116 kDa subunit a isoform 3 93 kDa 3 16 1 2 OS = Homosapiens OX = 9606 GN = TCIRG1 PE = 1 SV = 3 WD repeat-containing protein61 OS = Homo sapiens 34 kDa 3 0 1 4 OX = 9606 GN = WDR61 PE = 1 SV = 139S ribosomal protein L41, mitochondrial OS = Homo 15 kDa 2 0 0 5sapiens OX = 9606 GN = MRPL41 PE = 1 SV = 1 40S ribosomal protein S25 OS= Homo sapiens OX = 9606 14 kDa 2 0 8 7 GN = RPS25 PE = 1 SV = 1 60Sacidic ribosomal protein P1 OS = Homo sapiens 12 kDa 2 13 3 5 OX = 9606GN = RPLP1 PE = 1 SV = 1 Actin-like protein 6A OS = Homo sapiens OX =9606 47 kDa 2 12 0 2 GN = ACTL6A PE = 1 SV = 1 Actin-related protein 2/3complex subunit 5 OS = Homo 16 kDa 2 15 5 7 sapiens OX = 9606 GN = ARPC5PE = 1 SV = 3 Actin-related protein 2/3 complex subunit 5-like protein17 kDa 2 0 6 5 OS = Homo sapiens OX = 9606 GN = ARPC5L PE = 1 SV = 1Activated RNA polymerase II transcriptional coactivator 14 kDa 2 1 0 2p15 OS = Homo sapiens OX = 9606 GN = SUB1 PE = 1 SV = 3 Alcoholdehydrogenase [NADP(+)] OS = Homo sapiens 37 kDa 2 0 0 1 OX = 9606 GN =AKR1A1 PE = 1 SV = 3 Aspartyl/asparaginyl beta-hydroxylase OS = Homo 86kDa 2 4 1 3 sapiens OX = 9606 GN = ASPH PE = 1 SV = 3 ATP synthasesubunit f, mitochondrial OS = Homo 11 kDa 2 0 2 2 sapiens OX = 9606 GN =ATP5MF PE = 1 SV = 1 ATP-binding cassette sub-family B member 6, 78 kDa2 0 0 0 mitochondrial (Fragment) OS = Homo sapiens OX = 9606 GN = ABCB6PE = 1 SV = 1 Catenin delta-1 OS = Homo sapiens OX = 9606 108 kDa 2 13 01 GN = CTNND1 PE = 1 SV = 1 Cathepsin B OS = Homo sapiens OX = 9606 GN =CTSB 38 kDa 2 0 0 0 PE = 1 SV = 3 Caveolin-1 OS = Homo sapiens OX = 9606GN = CAV1 20 kDa 2 11 2 4 PE = 1 SV = 4 Cell cycle and apoptosisregulator protein 2 OS = Homo 103 kDa 2 1 0 4 sapiens GN = CCAR2 PE = 1SV = 2 Centromere protein V OS = Homo sapiens OX = 9606 30 kDa 2 3 0 1GN = CENPV PE = 1 SV = 1 Citrate synthase OS = Homo sapiens OX = 9606 GN= CS 50 kDa 2 0 0 3 PE = 1 SV = 1 Cytochrome b-c1 complex subunit 1,mitochondrial 53 kDa 2 0 0 1 OS = Homo sapiens OX = 9606 GN = UQCRC1 PE= 1 SV = 3 Cytochrome c oxidase subunit 7A2, mitochondrial 9 kDa 2 0 0 1OS = Homo sapiens OX = 9606 GN = COX7A2 PE = 1 SV = 1 Dystonin OS = Homosapiens OX = 9606 GN = DST PE = 1 861 kDa 2 0 0 0 SV = 4 EHdomain-containing protein 2 OS = Homo sapiens 61 kDa 2 2 0 3 OX = 9606GN = EHD2 PE = 1 SV = 2 Enhancer of mRNA-decapping protein 4 OS = Homo152 kDa 2 0 0 0 sapiens GN = EDC4 PE = 1 SV = 1 Eukaryotic translationinitiation factor 5B OS = Homo 139 kDa 2 0 0 2 sapiens OX = 9606 GN =EIF5B PE = 1 SV = 4 Exosome RNA helicase MTR4 OS = Homo sapiens 118 kDa2 0 0 1 OX = 9606 GN = MTREX PE = 1 SV = 3 Far upstream element-bindingprotein 2 OS = Homo 73 kDa 2 0 0 0 sapiens OX = 9606 GN = KHSRP PE = 1SV = 4 Ferritin heavy chain OS = Homo sapiens OX = 9606 21 kDa 2 0 0 1GN = FTH1 PE = 1 SV = 2 FH2 domain-containing protein 1 OS = Homosapiens 125 kDa 2 0 0 0 OX = 9606 GN = FHDC1 PE = 1 SV = 2Glutaredoxin-3 OS = Homo sapiens OX = 9606 37 kDa 2 0 0 0 GN = GLRX3 PE= 1 SV = 2 GTP-binding protein SAR1a OS = Homo sapiens 22 kDa 2 0 0 3 OX= 9606 GN = SAR1A PE = 1 SV = 1 Guanine nucleotide-binding proteinG(I)/G(S)/G(T) 37 kDa 2 14 0 0 subunit beta-2 OS = Homo sapiens OX =9606 GN = GNB2 PE = 1 SV = 3 Heterogeneous nuclear ribonucleoproteinU-like protein 2 85 kDa 2 8 0 2 OS = Homo sapiens OX = 9606 GN =HNRNPUL2 PE = 1 SV = 1 High mobility group protein B3 OS = Homo sapiens23 kDa 2 0 0 3 OX = 9606 GN = HMGB3 PE = 1 SV = 4 Inversin OS = Homosapiens OX = 9606 GN = INVS PE = 1 118 kDa 2 0 0 0 SV = 2 Isoform 10 ofCD44 antigen OS = Homo sapiens 53 kDa 2 27 5 2 OX = 9606 GN = CD44Isoform 2 of 40S ribosomal protein S20 OS = Homo 16 kDa 2 7 1 3 sapiensOX = 9606 GN = RPS20 Isoform 2 of AP-2 complex subunit alpha-2 OS = Homo104 kDa 2 8 0 3 sapiens OX = 9606 GN = AP2A2 Isoform 2 of ATP-bindingcassette sub-family F member 92 kDa 2 6 2 2 1 OS = Homo sapiens OX =9606 GN = ABCF1 Isoform 2 of cAMP-dependent protein kinase type II- 43kDa 2 11 0 3 alpha regulatory subunit OS = Homo sapiens OX = 9606 GN =PRKAR2A Isoform 2 of Chromatin target of PRMT1 protein 27 kDa 2 4 2 2 OS= Homo sapiens OX = 9606 GN = CHTOP Isoform 2 of Collagen alpha-1(XXII)chain OS = Homo 159 kDa 2 0 0 1 sapiens OX = 9606 GN = COL22A1 Isoform 2of DNA repair protein RAD50 OS = Homo 155 kDa 2 0 0 1 sapiens OX = 9606GN = RAD50 Isoform 2 of E3 ubiquitin-protein ligase RNF213 596 kDa 2 0 01 OS = Homo sapiens OX = 9606 GN = RNF213 Isoform 2 of Electron transferflavoprotein subunit alpha, 30 kDa 2 1 0 5 mitochondrial OS = Homosapiens OX = 9606 GN = ETFA Isoform 2 of Enoyl-CoA delta isomerase 2,mitochondrial 40 kDa 2 0 0 0 OS = Homo sapiens OX = 9606 GN = ECI2Isoform 2 of Eukaryotic peptide chain release factor 45 kDa 2 2 0 2subunit 1 OS = Homo sapiens OX = 9606 GN = ETF1 Isoform 2 of Eukaryotictranslation initiation factor 3 58 kDa 2 2 0 0 subunit D OS = Homosapiens OX = 9606 GN = EIF3D Isoform 2 of Fermitin family homolog 2 OS =Homo 72 kDa 2 0 0 6 sapiens OX = 9606 GN = FERMT2 Isoform 2 ofGlutamine--fructose-6-phosphate 77 kDa 2 5 0 1 aminotransferase[isomerizing] 1 OS = Homo sapiens OX = 9606 GN = GFPT1 Isoform 2 ofGuanine nucleotide-binding protein-like 3 61 kDa 2 3 0 0 OS = Homosapiens OX = 9606 GN = GNL3 Isoform 2 of Histidine--tRNA ligase,cytoplasmic 53 kDa 2 3 0 4 OS = Homo sapiens OX = 9606 GN = HARS Isoform2 of Histone H1.0 OS = Homo sapiens OX = 9606 19 kDa 2 1 3 3 GN = H1F0Isoform 2 of Interferon-induced, double-stranded RNA- 57 kDa 2 1 0 1activated protein kinase OS = Homo sapiens OX = 9606 GN = EIF2AK2Isoform 2 of Neurotrimin OS = Homo sapiens OX = 9606 38 kDa 2 0 0 0 GN =NTM Isoform 2 of Neutral cholesterol ester hydrolase 1 47 kDa 2 1 0 3 OS= Homo sapiens OX = 9606 GN = NCEH1 Isoform 2 of Nucleosome assemblyprotein 1-like 1 43 kDa 2 10 1 3 OS = Homo sapiens OX = 9606 GN = NAP1L1Isoform 2 of Nucleosome assembly protein 1-like 4 44 kDa 2 0 0 3 OS =Homo sapiens OX = 9606 GN = NAP1L4 Isoform 2 of Polyadenylate-bindingprotein 2 OS = Homo 31 kDa 2 7 0 2 sapiens OX = 9606 GN = PABPN1 Isoform2 of Pre-mRNA-splicing factor SYF2 OS = Homo 24 kDa 2 0 0 3 sapiens OX =9606 GN = SYF2 Isoform 2 of Procollagen-lysine,2-oxoglutarate 5- 88 kDa2 12 0 1 dioxygenase 1 OS = Homo sapiens OX = 9606 GN = PLOD1 Isoform 2of Procollagen-lysine,2-oxoglutarate 5- 87 kDa 2 2 0 0 dioxygenase 2 OS= Homo sapiens OX = 9606 GN = PLOD2 Isoform 2 of Protein enabled homologOS = Homo sapiens 64 kDa 2 26 1 18 OX = 9606 GN = ENAH Isoform 2 ofProtein SGT1 homolog OS = Homo sapiens 38 kDa 2 0 0 0 OX = 9606 GN =SUGT1 Isoform 2 of Protein transport protein Sec16A OS = Homo 229 kDa 20 0 1 sapiens OX = 9606 GN = SEC16A Isoform 2 of RNA-binding proteinwith serine-rich 32 kDa 2 3 1 2 domain 1 OS = Homo sapiens OX = 9606 GN= RNPS1 Isoform 2 of Signal recognition particle subunit SRP68 67 kDa 29 0 0 OS = Homo sapiens OX = 9606 GN = SRP68 Isoform 2 of Syntenin-1 OS= Homo sapiens OX = 9606 32 kDa 2 6 0 0 GN = SDCBP Isoform 2 ofTranslocating chain-associated membrane 40 kDa 2 6 4 5 protein 1 OS =Homo sapiens OX = 9606 GN = TRAM1 Isoform 2 of Trifunctional enzymesubunit beta, 49 kDa 2 51 9 10 mitochondrial OS = Homo sapiens OX = 9606GN = HADHB Isoform 2 of UDP-glucose: glycoprotein 175 kDa 2 2 0 1glucosyltransferase 1 OS = Homo sapiens OX = 9606 GN = UGGT1 Isoform 3of 28S ribosomal protein S29, mitochondrial 42 kDa 2 5 2 1 OS = Homosapiens GN = DAP3 Isoform 3 of Basic leucine zipper and W2 domain- 51kDa 2 1 0 4 containing protein 1 OS = Homo sapiens OX = 9606 GN = BZW1Isoform 3 of E3 ubiquitin-protein ligase HUWE1 481 kDa 2 4 0 0 OS = Homosapiens GN = HUWE1 Isoform 3 of Heterogeneous nuclear ribonucleoprotein32 kDa 2 8 0 3 H3 OS = Homo sapiens GN = HNRNPH3 Isoform 3 ofIntegrin-linked protein kinase OS = Homo 36 kDa 2 0 0 2 sapiens OX =9606 GN = ILK Isoform 3 of Protein virilizer homolog OS = Homo sapiens201 kDa 2 0 0 0 OX = 9606 GN = VIRMA Isoform 3 of Ubiquitin-associatedprotein 2-like 113 kDa 2 7 0 7 OS = Homo sapiens OX = 9606 GN = UBAP2LIsoform 4 of LIM domain and actin-binding protein 1 85 kDa 2 49 6 4 OS =Homo sapiens OX = 9606 GN = LIMA1 Isoform 4 of WASH complex subunit 2COS = Homo 145 kDa 2 0 0 0 sapiens OX = 9606 GN = WASHC2C Isoform Delta10 of Calcium/calmodulin-dependent 56 kDa 2 5 0 0 protein kinase type IIsubunit delta OS = Homo sapiens OX = 9606 GN = CAMK2D Isoform LAMP-2B ofLysosome-associated membrane 45 kDa 2 0 0 2 glycoprotein 2 OS = Homosapiens OX = 9606 GN = LAMP2 Leucine-rich repeat flightless-interactingprotein 1 89 kDa 2 0 0 1 OS = Homo sapiens OX = 9606 GN = LRRFIP1 PE = 1SV = 2 Lysophospholipid acyltransferase 7 OS = Homo sapiens 53 kDa 2 2 00 GN = MBOAT7 PE = 1 SV = 2 Microsomal glutathione S-transferase 3 OS =Homo 17 kDa 2 1 1 2 sapiens OX = 9606 GN = MGST3 PE = 1 SV = 1Mitochondrial import receptor subunit TOM40 homolog 38 kDa 2 0 0 0 OS =Homo sapiens OX = 9606 GN = TOMM40 PE = 1 SV = 1 Mucin-19 OS = Homosapiens OX = 9606 GN = MUC19 805 kDa 2 0 0 0 PE = 1 SV = 3 NADHdehydrogenase [ubiquinone] iron-sulfur protein 3, 30 kDa 2 4 0 4mitochondrial OS = Homo sapiens OX = 9606 GN = NDUFS3 PE = 1 SV = 1Nascent polypeptide-associated complex subunit alpha, 205 kDa 2 6 0 4muscle-specific form OS = Homo sapiens OX = 9606 GN = NACA PE = 1 SV = 1Nicotinamide phosphoribosyltransferase OS = Homo 56 kDa 2 2 0 3 sapiensOX = 9606 GN = NAMPT PE = 1 SV = 1 Non-histone chromosomal proteinHMG-14 OS = Homo 12 kDa 2 11 3 6 sapiens OX = 9606 GN = HMGN1 PE = 1 SV= 1 Nucleolar protein 11 OS = Homo sapiens OX = 9606 81 kDa 2 0 0 0 GN =NOL11 PE = 1 SV = 1 Obg-like ATPase 1 OS = Homo sapiens OX = 9606 45 kDa2 4 0 3 GN = OLA1 PE = 1 SV = 2 PDZ and LIM domain protein 5 OS = Homosapiens 64 kDa 2 3 0 2 GN = PDLIM5 PE = 1 SV = 5 Peroxiredoxin-2 OS =Homo sapiens OX = 9606 22 kDa 2 7 0 1 GN = PRDX2 PE = 1 SV = 5Phospholipid-transporting ATPase IB OS = Homo sapiens 129 kDa 2 0 2 0 OX= 9606 GN = ATP8A2 PE = 1 SV = 3 Phosphoserine aminotransferase OS =Homo sapiens 40 kDa 2 0 0 0 OX = 9606 GN = PSAT1 PE = 1 SV = 2Plasminogen activator inhibitor 1 OS = Homo sapiens 45 kDa 2 90 41 29 OX= 9606 GN = SERPINE1 PE = 1 SV = 1 pre-rRNA processing protein FTSJ3 OS= Homo sapiens 97 kDa 2 3 0 1 OX = 9606 GN = FTSJ3 PE = 1 SV = 2 PRKCapoptosis WT1 regulator protein OS = Homo 37 kDa 2 0 1 6 sapiens OX =9606 GN = PAWR PE = 1 SV = 1 Proteasome activator complex subunit 2 OS =Homo 27 kDa 2 0 0 2 sapiens OX = 9606 GN = PSME2 PE = 1 SV = 4Proteasome subunit alpha type-5 OS = Homo sapiens 26 kDa 2 2 0 2 OX =9606 GN = PSMA5 PE = 1 SV = 3 Proteasome subunit alpha type-7 OS = Homosapiens 28 kDa 2 7 0 2 OX = 9606 GN = PSMA7 PE = 1 SV = 1 Proteasomesubunit beta type-2 OS = Homo sapiens 23 kDa 2 0 0 4 OX = 9606 GN =PSMB2 PE = 1 SV = 1 Protein flightless-1 homolog OS = Homo sapiens 145kDa 2 0 0 1 OX = 9606 GN = FLII PE = 1 SV = 2 Protein SEC13 homolog OS =Homo sapiens OX = 9606 36 kDa 2 4 0 4 GN = SEC13 PE = 1 SV = 3Ras-related protein Rab-5C OS = Homo sapiens OX = 9606 23 kDa 2 7 0 3 GN= RAB5C PE = 1 SV = 2 Ras-related protein Rab-7a OS = Homo sapiens OX =9606 23 kDa 2 0 0 2 GN = RAB7A PE = 1 SV = 1 Remodeling and spacingfactor 1 OS = Homo sapiens 164 kDa 2 9 0 0 GN = RSF1 PE = 1 SV = 2Serine beta-lactamase-like protein LACTB, 61 kDa 2 13 5 3 mitochondrialOS = Homo sapiens OX = 9606 GN = LACTB PE = 1 SV = 2Serine/arginine-rich splicing factor 1 OS = Homo sapiens 28 kDa 2 7 0 2GN = SRSF1 PE = 1 SV = 2 Structural maintenance of chromosomes protein1A 143 kDa 2 7 0 0 OS = Homo sapiens OX = 9606 GN = SMC1A PE = 1 SV = 2SWISS-PROT: P02777 (Bos taurus) similar to Platelet 24 kDa 2 3 3 3factor 4 SWISS-PROT: Q2UVX4 (Bos taurus) Complement C3 187 kDa 2 7 6 4precursor SWISS-PROT: Q95121 (Bos taurus) Pigment epithelium- 46 kDa 2 03 3 derived factor precursor Transaldolase OS = Homo sapiens OX = 960638 kDa 2 0 0 3 GN = TALDO1 PE = 1 SV = 2 Translationally-controlledtumor protein OS = Homo 20 kDa 2 0 0 2 sapiens OX = 9606 GN = TPT1 PE =1 SV = 1 Translocon-associated protein subunit alpha OS = Homo 32 kDa 25 3 5 sapiens OX = 9606 GN = SSR1 PE = 1 SV = 3 Transmembrane emp24domain-containing protein 10 25 kDa 2 0 6 8 OS = Homo sapiens OX = 9606GN = TMED10 PE = 1 SV = 2 Transmembrane emp24 domain-containing protein9 27 kDa 2 0 1 2 OS = Homo sapiens OX = 9606 GN = TMED9 PE = 1 SV = 2TREMBL: Q1RMN8 (Bos taurus) Similar to 25 kDa 2 0 0 0 Immunoglobulinlambda-like polypeptide 1 Tripeptidyl-peptidase 1 OS = Homo sapiens GN =TPP1 61 kDa 2 0 2 3 PE = 1 SV = 2 1,4-alpha-glucan-branching enzyme OS =Homo sapiens 80 kDa 1 0 0 3 OX = 9606 GN = GBE1 PE = 1 SV = 31-phosphatidylinositol 4,5-bisphosphate 139 kDa 1 0 0 2phosphodiesterase beta-3 OS = Homo sapiens GN = PLCB3 PE = 1 SV = 2 28Sribosomal protein S35, mitochondrial OS = Homo 37 kDa 1 2 0 2 sapiens GN= MRPS35 PE = 1 SV = 1 40S ribosomal protein S30 OS = Homo sapiens OX =9606 7 kDa 1 0 3 2 GN = FAU PE = 1 SV = 1 60S ribosomal protein L11 OS =Homo sapiens OX = 9606 20 kDa 1 18 1 3 GN = RPL11 PE = 1 SV = 2 60Sribosomal protein L35a OS = Homo sapiens 13 kDa 1 0 2 2 OX = 9606 GN =RPL35A PE = 1 SV = 2 60S ribosomal protein L37a OS = Homo sapiens 10 kDa1 0 5 5 OX = 9606 GN = RPL37A PE = 1 SV = 2 Alpha-parvin OS = Homosapiens OX = 9606 GN = PARVA 42 kDa 1 3 0 4 PE = 1 SV = 1 AT-richinteractive domain-containing protein 1A 242 kDa 1 3 0 0 OS = Homosapiens OX = 9606 GN = ARID1 A PE = 1 SV = 3 Basigin OS = Homo sapiensOX = 9606 GN = BSG PE = 1 42 kDa 1 3 0 1 SV = 2 Biorientation ofchromosomes in cell division protein 1- 330 kDa 1 0 0 3 like 1 OS = Homosapiens OX = 9606 GN = BOD1L1 PE = 1 SV = 2 Cell division cycle 5-likeprotein OS = Homo sapiens 92 kDa 1 3 0 4 OX = 9606 GN = CDC5L PE = 1 SV= 2 Coactosin-like protein OS = Homo sapiens OX = 9606 16 kDa 1 0 0 2 GN= COTL1 PE = 1 SV = 3 Collagen alpha-1(III) chain OS = Homo sapiens OX =9606 139 kDa 1 2 0 1 GN = COL3A1 PE = 1 SV = 4 Collagen alpha-2(I) chainOS = Homo sapiens OX = 9606 129 kDa 1 12 7 3 GN = COL1A2 PE = 1 SV = 7Collagen alpha-2(V) chain OS = Homo sapiens OX = 9606 145 kDa 1 0 15 6GN = COL5A2 PE = 1 SV = 3 Collagen alpha-6(VI) chain OS = Homo sapiensOX = 9606 247 kDa 1 0 3 2 GN = COL6A6 PE = 1 SV = 2 Complement component1 Q subcomponent-binding 31 kDa 1 0 0 4 protein, mitochondrial OS = Homosapiens OX = 9606 GN = C1QBP PE = 1 SV = 1 Cysteine and glycine-richprotein 1 OS = Homo sapiens 21 kDa 1 0 0 2 OX = 9606 GN = CSRP1 PE = 1SV = 3 Cytochrome b-c1 complex subunit 9 OS = Homo sapiens 7 kDa 1 2 0 2OX = 9606 GN = UQCR10 PE = 1 SV = 3 Cytochrome c1, heme protein,mitochondrial OS = Homo 35 kDa 1 0 0 3 sapiens OX = 9606 GN = CYC1 PE =1 SV = 3 DDRGK domain-containing protein 1 OS = Homo sapiens 36 kDa 1 01 2 OX = 9606 GN = DDRGK1 PE = 1 SV = 2 Destrin OS = Homo sapiens OX =9606 GN = DSTN PE = 1 19 kDa 1 3 0 6 SV = 3 Dihydrolipoyl dehydrogenase,mitochondrial OS = Homo 54 kDa 1 8 0 2 sapiens OX = 9606 GN = DLD PE = 1SV = 2 DNA-directed RNA polymerases I, II, and III subunit 25 kDa 1 0 51 RPABC1 OS = Homo sapiens OX = 9606 GN = POLR2E PE = 1 SV = 4 ELAV-likeprotein 1 OS = Homo sapiens OX = 9606 36 kDa 1 3 0 1 GN = ELAVL1 PE = 1SV = 2 Eukaryotic translation initiation factor 3 subunit G 36 kDa 1 0 03 OS = Homo sapiens OX = 9606 GN = EIF3G PE = 1 SV = 2 Farnesylpyrophosphate synthase OS = Homo sapiens 48 kDa 1 3 0 1 OX = 9606 GN =FDPS PE = 1 SV = 4 Fibrillin-1 OS = Homo sapiens OX = 9606 GN = FBN1 312kDa 1 0 2 0 PE = 1 SV = 3 Fumarate hydratase, mitochondrial OS = Homosapiens 55 kDa 1 0 0 2 GN = FH PE = 1 SV = 3 Growth/differentiationfactor 15 OS = Homo sapiens 34 kDa 1 0 7 6 OX = 9606 GN = GDF15 PE = 1SV = 3 Histone deacetylase complex subunit SAP18 OS = Homo 20 kDa 1 0 02 sapiens OX = 9606 GN = SAP18 PE = 1 SV = 1 Histone H1x OS = Homosapiens OX = 9606 GN = H1FX 22 kDa 1 0 0 2 PE = 1 SV = 1Hydrocephalus-inducing protein homolog OS = Homo 576 kDa 1 0 1 0 sapiensOX = 9606 GN = HYDIN PE = 1 SV = 3 Insulin-like growth factor 2mRNA-binding protein 3 64 kDa 1 18 2 0 OS = Homo sapiens GN = IGF2BP3 PE= 1 SV = 2 Isoform 2 of Adipocyte plasma membrane-associated 32 kDa 1 00 3 protein OS = Homo sapiens OX = 9606 GN = APMAP Isoform 2 ofAspartate aminotransferase, mitochondrial 43 kDa 1 0 0 3 OS = Homosapiens OX = 9606 GN = GOT2 Isoform 2 of BH3-interacting domain deathagonist 27 kDa 1 4 0 0 OS = Homo sapiens OX = 9606 GN = BID Isoform 2 ofDnaJ homolog subfamily A member 3, 50 kDa 1 0 0 3 mitochondrial OS =Homo sapiens OX = 9606 GN = DNAJA3 Isoform 2 of Glia-derived nexin OS =Homo sapiens 44 kDa 1 8 0 0 OX = 9606 GN = SERPINE2 Isoform 2 of Histonedeacetylase 2 OS = Homo sapiens 52 kDa 1 4 0 1 OX = 9606 GN = HDAC2Isoform 2 of Lysosomal protective protein OS = Homo 52 kDa 1 1 0 2sapiens OX = 9606 GN = CTSA Isoform 2 of Lysosome membrane protein 2 OS= Homo 38 kDa 1 4 0 0 sapiens OX = 9606 GN = SCARB2 Isoform 2 of Myosinphosphatase Rho-interacting protein 118 kDa 1 13 0 0 OS = Homo sapiensOX = 9606 GN = MPRIP Isoform 2 of NADH dehydrogenase [ubiquinone] 1alpha 25 kDa 1 13 1 3 subcomplex subunit 13 OS = Homo sapiens OX = 9606GN = NDUFA13 Isoform 2 of NADH dehydrogenase [ubiquinone] iron- 52 kDa 15 0 1 sulfur protein 2, mitochondrial OS = Homo sapiens OX = 9606 GN =NDUFS2 Isoform 2 of Nesprin-2 OS = Homo sapiens OX = 9606 799 kDa 1 12 10 GN = SYNE2 Isoform 2 of Nuclear pore complex protein Nup214 213 kDa 10 0 3 OS = Homo sapiens OX = 9606 GN = NUP214 Isoform 2 ofPhenylalanine--tRNA ligase alpha subunit 54 kDa 1 7 1 4 OS = Homosapiens OX = 9606 GN = FARSA Isoform 2 of Probable 28S rRNA(cytosine(4447)-C(5))- 89 kDa 1 6 0 6 methyltransferase OS = Homosapiens OX = 9606 GN = NOP2 Isoform 2 of Protein FAM98A OS = Homosapiens 55 kDa 1 2 0 3 OX = 9606 GN = FAM98A Isoform 2 of Protein TFG OS= Homo sapiens OX = 9606 43 kDa 1 4 0 4 GN = TFG Isoform 2 of Pyruvatedehydrogenase E1 component 37 kDa 1 7 0 3 subunit beta, mitochondrial OS= Homo sapiens OX = 9606 GN = PDHB Isoform 2 of UTP--glucose-1-phosphate56 kDa 1 3 0 7 uridylyltransferase OS = Homo sapiens OX = 9606 GN = UGP2Isoform 2 of V-type proton ATPase 116 kDa subunit a 96 kDa 1 11 0 3isoform 1 OS = Homo sapiens OX = 9606 GN = ATP6V0A1 Isoform 2B ofCytoplasmic dynein 1 intermediate chain 2 71 kDa 1 16 1 3 OS = Homosapiens OX = 9606 GN = DYNC1I2 Isoform 3 of Activating signalcointegrator 1 complex 77 kDa 1 6 0 1 subunit 2 OS = Homo sapiens OX =9606 GN = ASCC2 Isoform 3 of Apoptosis-inducing factor 1, mitochondrial66 kDa 1 3 0 4 OS = Homo sapiens OX = 9606 GN = AIFM1 Isoform 3 ofCalumenin OS = Homo sapiens OX = 9606 38 kDa 1 1 0 2 GN = CALU Isoform 3of Fragile X mental retardation syndrome- 60 kDa 1 4 0 1 related protein1 OS = Homo sapiens OX = 9606 GN = FXR1 Isoform 3 of GRB10-interactingGYF protein 2 149 kDa 1 0 0 0 OS = Homo sapiens OX = 9606 GN = GIGYF2Isoform 3 of Pyrroline-5-carboxylate reductase 1, 36 kDa 1 8 0 0mitochondrial OS = Homo sapiens OX = 9606 GN = PYCR1 Isoform 3 ofRas-related protein R-Ras2 OS = Homo 20 kDa 1 5 0 1 sapiens GN = RRAS2Isoform 3 of Signal peptidase complex catalytic subunit 21 kDa 1 2 1 3SEC11A OS = Homo sapiens OX = 9606 GN = SEC11A Isoform 4 of Cadherin-13OS = Homo sapiens OX = 9606 83 kDa 1 23 6 2 GN = CDH13 Isoform 4 ofDipeptidyl peptidase 3 OS = Homo sapiens 79 kDa 1 0 0 2 OX = 9606 GN =DPP3 Isoform 4 of Dynamin-1-like protein OS = Homo sapiens 81 kDa 1 2 02 OX = 9606 GN = DNM1L Isoform 4 of Dynamin-2 OS = Homo sapiens OX =9606 98 kDa 1 13 0 6 GN = DNM2 Isoform 4 of Nexilin OS = Homo sapiens OX= 9606 73 kDa 1 28 12 7 GN = NEXN Isoform 5 of Obscurin OS = Homosapiens OX = 9606 925 kDa 1 0 0 1 GN = OBSCN Isoform 6 of RNA-bindingprotein EWS OS = Homo 63 kDa 1 6 0 6 sapiens OX = 9606 GN = EWSR1Isoform Heart of ATP synthase subunit gamma, 33 kDa 1 6 0 4mitochondrial OS = Homo sapiens OX = 9606 GN = ATP5F1C IsoformMitochondrial of Lysine--tRNA ligase 71 kDa 1 4 0 1 OS = Homo sapiens OX= 9606 GN = KARS Leucine-rich repeat-containing protein 17 OS = Homo 52kDa 1 11 9 4 sapiens OX = 9606 GN = LRRC17 PE = 2 SV = 1 Leucyl-cystinylaminopeptidase OS = Homo sapiens 117 kDa 1 4 3 2 OX = 9606 GN = LNPEP PE= 1 SV = 3 LIM and SH3 domain protein 1 OS = Homo sapiens 30 kDa 1 1 0 3OX = 9606 GN = LASP1 PE = 1 SV = 2 Matrix metalloproteinase-14 OS = Homosapiens 66 kDa 1 0 0 2 OX = 9606 GN = MMP14 PE = 1 SV = 3Microtubule-associated protein RP/EB family member 1 30 kDa 1 4 0 4 OS =Homo sapiens OX = 9606 GN = MAPRE1 PE = 1 SV = 3 Mitochondrial2-oxoglutarate/malate carrier protein 34 kDa 1 7 0 1 OS = Homo sapiensOX = 9606 GN = SLC25A11 PE = 1 SV = 3 Mitochondrial fission 1 protein OS= Homo sapiens 17 kDa 1 0 0 3 OX = 9606 GN = FIS1 PE = 1 SV = 2 NADHdehydrogenase [ubiquinone] 1 alpha subcomplex 17 kDa 1 6 1 2 subunit 12OS = Homo sapiens OX = 9606 GN = NDUFA12 PE = 1 SV = 1 PDZdomain-containing protein 4 OS = Homo sapiens 86 kDa 1 0 0 0 OX = 9606GN = PDZD4 PE = 1 SV = 1 Peroxisomal membrane protein PEX14 OS = Homo 41kDa 1 3 0 2 sapiens OX = 9606 GN = PEX14 PE = 1 SV = 1Platelet-activating factor acetylhydrolase IB subunit beta 26 kDa 1 6 01 OS = Homo sapiens GN = PAFAH1B2 PE = 1 SV = 1 Podocalyxin OS = Homosapiens GN = PODXL PE = 1 59 kDa 1 0 0 2 SV = 2 Prolyl 3-hydroxylase 3OS = Homo sapiens OX = 9606 82 kDa 1 0 0 2 GN = P3H3 PE = 1 SV = 1Prolyl 4-hydroxylase subunit alpha-2 OS = Homo sapiens 61 kDa 1 2 0 0 OX= 9606 GN = P4HA2 PE = 1 SV = 1 Proteasome subunit alpha type-6 OS =Homo sapiens 27 kDa 1 8 0 1 OX = 9606 GN = PSMA6 PE = 1 SV = 1Proteasome subunit beta type-5 OS = Homo sapiens 28 kDa 1 2 0 0 GN =PSMB5 PE = 1 SV = 3 Protein CYR61 OS = Homo sapiens OX = 9606 42 kDa 1 027 24 GN = CYR61 PE = 1 SV = 1 Protein mago nashi homolog OS = Homosapiens 17 kDa 1 7 0 0 OX = 9606 GN = MAGOH PE = 1 SV = 1 RNA-bindingprotein 14 OS = Homo sapiens OX = 9606 69 kDa 1 18 2 5 GN = RBM14 PE = 1SV = 2 RuvB-like 1 OS = Homo sapiens OX = 9606 GN = RUVBL1 50 kDa 1 18 02 PE = 1 SV = 1 Splicing factor 3B subunit 4 OS = Homo sapiens 44 kDa 10 0 2 OX = 9606 GN = SF3B4 PE = 1 SV = 1 Splicing factor U2AF 26 kDasubunit OS = Homo sapiens 4 kDa 1 0 2 1 OX = 9606 GN = U2AF1L4 PE = 4 SV= 1 SWISS-PROT: P04258 (Bos taurus) Similar to Collagen 138 kDa 1 9 0 0alpha 1(III) chain TREMBL: A2I7N3; Q27984 (Bos taurus) SERPINA3-7 47 kDa1 5 1 1 Unconventional myosin-Ie OS = Homo sapiens OX = 9606 127 kDa 120 5 4 GN = MYO1E PE = 1 SV = 2 Vacuolar protein sorting-associatedprotein VTA1 34 kDa 1 0 0 2 homolog OS = Homo sapiens OX = 9606 GN =VTA1 PE = 1 SV = 1 Very-long-chain (3R)-3-hydroxyacyl-CoA dehydratase 343 kDa 1 8 1 2 OS = Homo sapiens GN = HACD3 PE = 1 SV = 2 von Willebrandfactor A domain-containing protein 1 47 kDa 1 7 0 0 OS = Homo sapiens OX= 9606 GN = VWA1 PE = 1 SV = 1 (Bos taurus) 63 kDa protein 63 kDa 0 55 00 10 kDa heat shock protein, mitochondrial OS = Homo 11 kDa 0 6 0 0sapiens GN = HSPE1 PE = 1 SV = 2 1-phosphatidylinositol 3-phosphate5-kinase OS = Homo 237 kDa 0 0 0 2 sapiens OX = 9606 GN = PIKFYVE PE = 1SV = 3 26S protease regulatory subunit 10B OS = Homo sapiens 46 kDa 0 90 0 GN = PSMC6 PE = 1 SV = 1 26S protease regulatory subunit 6A OS =Homo sapiens 49 kDa 0 30 0 0 GN = PSMC3 PE = 1 SV = 3 26S proteasomenon-ATPase regulatory subunit 14 35 kDa 0 17 0 0 OS = Homo sapiens GN =PSMD14 PE = 1 SV = 1 26S proteasome non-ATPase regulatory subunit 8 33kDa 0 3 0 0 OS = Homo sapiens GN = PSMD8 PE = 1 SV = 1 28S ribosomalprotein S34, mitochondrial OS = Homo 26 kDa 0 2 0 0 sapiens GN = MRPS34PE = 1 SV = 2 39S ribosomal protein L11, mitochondrial OS = Homo 21 kDa0 7 0 1 sapiens GN = MRPL11 PE = 1 SV = 1 39S ribosomal protein L34,mitochondrial OS = Homo 20 kDa 0 8 0 0 sapiens GN = MRPL34 PE = 1 SV = 13-ketoacyl-CoA thiolase, mitochondrial OS = Homo 42 kDa 0 0 0 2 sapiensOX = 9606 GN = ACAA2 PE = 1 SV = 2 40S ribosomal protein S13 OS = Homosapiens OX = 9606 17 kDa 0 85 19 19 GN = RPS13 PE = 1 SV = 2 40Sribosomal protein S15a OS = Homo sapiens 11 kDa 0 25 0 0 GN = RPS15A PE= 1 SV = 1 40S ribosomal protein S18 OS = Homo sapiens 18 kDa 0 51 0 0GN = RPS18 PE = 1 SV = 3 40S ribosomal protein S28 OS = Homo sapiens 8kDa 0 6 0 0 GN = RPS28 PE = 1 SV = 1 40S ribosomal protein S29 OS = Homosapiens OX = 9606 7 kDa 0 16 1 1 GN = RPS29 PE = 1 SV = 2 40S ribosomalprotein S3a OS = Homo sapiens 30 kDa 0 53 0 0 GN = RPS3A PE = 1 SV = 240S ribosomal protein S4, Y isoform 1 OS = Homo 29 kDa 0 0 0 9 sapiensOX = 9606 GN = RPS4Y1 PE = 1 SV = 2 40S ribosomal protein SA (Fragment)OS = Homo sapiens 29 kDa 0 8 0 0 GN = RPSA PE = 1 SV = 8 60S acidicribosomal protein P2 OS = Homo sapiens 12 kDa 0 50 0 0 GN = RPLP2 PE = 1SV = 1 60S ribosomal protein L13a OS = Homo sapiens 24 kDa 0 33 0 0 GN =RPL13A PE = 1 SV = 2 60S ribosomal protein L14 OS = Homo sapiens 23 kDa0 34 0 0 GN = RPL14 PE = 1 SV = 4 60S ribosomal protein L18a OS = Homosapiens 21 kDa 0 31 0 0 GN = RPL18A PE = 1 SV = 2 60S ribosomal proteinL23a (Fragment) OS = Homo 18 kDa 0 32 0 0 sapiens GN = RPL23A PE = 1 SV= 1 60S ribosomal protein L29 OS = Homo sapiens 18 kDa 0 6 0 0 GN =RPL29 PE = 1 SV = 2 60S ribosomal protein L32 (Fragment) OS = Homo 16kDa 0 20 0 0 sapiens GN = RPL32 PE = 1 SV = 1 60S ribosomal protein L34OS = Homo sapiens OX = 9606 13 kDa 0 0 3 3 GN = RPL34 PE = 1 SV = 3 60Sribosomal protein L35 OS = Homo sapiens 15 kDa 0 24 0 0 GN = RPL35 PE =1 SV = 2 60S ribosomal protein L38 OS = Homo sapiens OX = 9606 8 kDa 0 05 4 GN = RPL38 PE = 1 SV = 2 78 kDa glucose-regulated protein OS = Homosapiens 72 kDa 0 188 0 0 GN = HSPA5 PE = 1 SV = 2 A disintegrin andmetalloproteinase with thrombospondin 105 kDa 0 12 3 0 motifs 1 OS =Homo sapiens OX = 9606 GN = ADAMTS1 PE = 1 SV = 4 Actin-related protein10 OS = Homo sapiens 46 kDa 0 3 0 0 GN = ACTR10 PE = 1 SV = 1Actin-related protein 2/3 complex subunit 1B OS = Homo 41 kDa 0 28 0 0sapiens GN = ARPC1B PE = 1 SV = 3 ADP-ribosyl cyclase/cyclic ADP-ribosehydrolase 2 36 kDa 0 3 5 2 OS = Homo sapiens OX = 9606 GN = BST1 PE = 1SV = 2 Aminopeptidase N OS = Homo sapiens GN = ANPEP 110 kDa 0 194 0 0PE = 1 SV = 4 Angiopoietin-related protein 4 OS = Homo sapiens 45 kDa 01 3 0 OX = 9606 GN = ANGPTL4 PE = 1 SV = 2 Annexin A3 OS = Homo sapiensOX = 9606 GN = ANXA3 36 kDa 0 0 0 3 PE = 1 SV = 3 Annexin A4 OS = Homosapiens OX = 9606 GN = ANXA4 36 kDa 0 0 0 2 PE = 1 SV = 4 AP-2 complexsubunit sigma OS = Homo sapiens 19 kDa 0 3 0 0 GN = AP2S1 PE = 1 SV = 1Arf-GAP with Rho-GAP domain, ANK repeat and PH 170 kDa 0 0 0 2domain-containing protein 3 OS = Homo sapiens OX = 9606 GN = ARAP3 PE =1 SV = 1 ATP synthase subunit epsilon-like protein, mitochondrial 6 kDa0 3 0 0 OS = Homo sapiens GN = ATP5EP2 PE = 3 SV = 1 ATP synthasesubunit g, mitochondrial OS = Homo 11 kDa 0 6 0 0 sapiens GN = ATP5L PE= 1 SV = 3 ATPase ASNA1 OS = Homo sapiens GN = ASNA1PE = 1 39 kDa 0 2 00 SV = 2 ATPase, H+ transporting, lysosomal accessory protein 1, 32 kDa0 7 0 0 isoform CRA_c OS = Homo sapiens GN = ATP6AP1 PE = 1 SV = 1ATP-dependent DNA helicase Q1 OS = Homo sapiens 73 kDa 0 2 0 0 GN =RECQL PE = 1 SV = 3 Barrier-to-autointegration factor OS = Homo sapiens10 kDa 0 24 0 0 GN = BANF1 PE = 1 SV = 1 Basement membrane-specificheparan sulfate 469 kDa 0 4914 0 0 proteoglycan core protein OS = Homosapiens GN = HSPG2 PE = 1 SV = 4 Beta-catenin-like protein 1 OS = Homosapiens 65 kDa 0 2 0 0 GN = CTNNBL1 PE = 1 SV = 1 Beta-galactosidase OS= Homo sapiens GN = GLB1 PE = 1 76 kDa 0 3 0 0 SV = 2 Biglycan OS = Homosapiens OX = 9606 GN = BGN PE = 1 42 kDa 0 0 23 0 SV = 2 Bonemorphogenetic protein 1 OS = Homo sapiens 111 kDa 0 14 3 0 OX = 9606 GN= BMP1 PE = 1 SV = 2 Brain acid soluble protein 1 OS = Homo sapiens 23kDa 0 10 0 0 GN = BASP1 PE = 1 SV = 2 C-1-tetrahydrofolate synthase,cytoplasmic OS = Homo 102 kDa 0 2 0 0 sapiens GN = MTHFD1 PE = 1 SV = 3Calmodulin OS = Homo sapiens GN = CALM1 PE = 1 17 kDa 0 24 0 0 SV = 2Calpain small subunit 1 OS = Homo sapiens 34 kDa 0 34 0 0 GN = CAPNS1 PE= 1 SV = 1 Carboxypeptidase M OS = Homo sapiens GN = CPM PE = 1 51 kDa 05 0 0 SV = 2 Cardiomyopathy-associated protein 5 OS = Homo sapiens 449kDa 0 2 0 0 GN = CMYA5 PE = 1 SV = 3 Casein kinase II subunit alpha OS =Homo sapiens 45 kDa 0 10 1 1 OX = 9606 GN = CSNK2A1 PE = 1 SV = 1CD2-associated protein OS = Homo sapiens GN = CD2AP 71 kDa 0 9 0 0 PE =1 SV = 1 CD59 glycoprotein OS = Homo sapiens OX = 9606 14 kDa 0 0 3 0 GN= CD59 PE = 1 SV = 1 Chloride intracellular channel protein 4 OS = Homo29 kDa 0 2 0 0 sapiens GN = CLIC4 PE = 1 SV = 4 Chloride intracellularchannel protein 6 OS = Homo 73 kDa 0 2 0 0 sapiens GN = CLIC6 PE = 2 SV= 3 Chromobox protein homolog 1 (Fragment) OS = Homo 19 kDa 0 5 0 0sapiens GN = CBX1 PE = 1 SV = 1 Coiled-coil domain-containing protein124 OS = Homo 26 kDa 0 6 0 0 sapiens GN = CCDC124 PE = 1 SV = 1Coiled-coil domain-containing protein 30 OS = Homo 91 kDa 0 2 0 0sapiens GN = CCDC30 PE = 2 SV = 1 Coiled-coil-helix-coiled-coil-helixdomain-containing 13 kDa 0 2 0 0 protein 1 OS = Homo sapiens GN = CHCHD1PE = 1 SV = 1 Collagen alpha-1(II) chain OS = Homo sapiens OX = 9606 142kDa 0 2 1 1 GN = COL2A1 PE = 1 SV = 3 Collagen alpha-1(VI) chain OS =Homo sapiens 108 kDa 0 3 0 0 GN = COL6A1 PE = 1 SV = 1 Collagenalpha-1(X) chain OS = Homo sapiens 66 kDa 0 3 0 0 GN = COL10A1 PE = 1 SV= 2 Collagen alpha-1(XXIII) chain OS = Homo sapiens 52 kDa 0 2 0 0 GN =COL23A1 PE = 1 SV = 1 Collagen alpha-1(XXVII) chain OS = Homo sapiens187 kDa 0 1 0 2 OX = 9606 GN = COL27A1 PE = 1 SV = 1 Collagenalpha-2(IX) chain OS = Homo sapiens OX = 9606 65 kDa 0 0 0 0 GN = COL9A2PE = 1 SV = 2 Collagen triple helix repeat-containing protein 1 26 kDa 03 1 1 OS = Homo sapiens OX = 9606 GN = CTHRC1 PE = 1 SV = 1 Connectivetissue growth factor OS = Homo sapiens 38 kDa 0 4 6 2 OX = 9606 GN =CTGF PE = 1 SV = 2 COP9 signalosome complex subunit 5 OS = Homo sapiens38 kDa 0 2 0 0 GN = COPS5 PE = 1 SV = 4 Coronin-1B OS = Homo sapiens GN= CORO1B PE = 1 54 kDa 0 9 0 0 SV = 1 Cullin-1 OS = Homo sapiens GN =CUL1 PE = 1 SV = 2 90 kDa 0 3 0 0 Cytochrome c (Fragment) OS = Homosapiens GN = CYCS 11 kDa 0 3 0 0 PE = 1 SV = 1 Cytochrome c oxidasesubunit 4 isoform 1, mitochondrial 20 kDa 0 5 0 0 OS = Homo sapiens GN =COX4I1 PE = 1 SV = 1 Cytoplasmic aconitate hydratase OS = Homo sapiens98 kDa 0 0 0 2 OX = 9606 GN = ACO1 PE = 1 SV = 3 Cytosolic non-specificdipeptidase OS = Homo sapiens 53 kDa 0 10 0 8 OX = 9606 GN = CNDP2 PE =1 SV = 2 Death-associated protein kinase 3 OS = Homo sapiens 53 kDa 0 60 0 OX = 9606 GN = DAPK3 PE = 1 SV = 1 Deoxyribose-phosphate aldolase OS= Homo sapiens 35 kDa 0 0 0 3 OX = 9606 GN = DERA PE = 1 SV = 2Desmoglein-1 OS = Homo sapiens GN = DSG1 PE = 1 114 kDa 0 6 0 0 SV = 2DNA damage-binding protein 1 OS = Homo sapiens 127 kDa 0 14 0 3 OX =9606 GN = DDB1 PE = 1 SV = 1 DNA topoisomerase 1 OS = Homo sapiens GN =TOP1 91 kDa 0 10 0 0 PE = 1 SV = 2 DNA-directed RNA polymerase IIsubunit RPB1 217 kDa 0 11 0 0 OS = Homo sapiens OX = 9606 GN = POLR2A PE= 1 SV = 2 DNA-directed RNA polymerase II subunit RPB2 134 kDa 0 14 0 0OS = Homo sapiens GN = POLR2B PE = 1 SV = 1 DNA-directed RNA polymeraseII subunit RPB3 31 kDa 0 9 0 0 OS = Homo sapiens GN = POLR2C PE = 1 SV =2 DNA-directed RNA polymerase II subunit RPB4 13 kDa 0 6 0 0 OS = Homosapiens GN = POLR2D PE = 1 SV = 1 DNA-directed RNA polymerases I, II,and III subunit 17 kDa 0 2 3 0 RPABC3 OS = Homo sapiens OX = 9606 GN =POLR2H PE = 1 SV = 4 DnaJ homolog subfamily B member 11 OS = Homo 41 kDa0 0 0 4 sapiens OX = 9606 GN = DNAJB11 PE = 1 SV = 1 Dolichol-phosphatemannosyltransferase subunit 1 30 kDa 0 2 0 0 OS = Homo sapiens GN = DPM1PE = 1 SV = 1 Dolichyl-diphosphooligosaccharide--protein 94 kDa 0 2 0 0glycosyltransferase subunit STT3B OS = Homo sapiens GN = STT3B PE = 1 SV= 1 Doublecortin domain-containing protein 2 OS = Homo 53 kDa 0 17 0 2sapiens OX = 9606 GN = DCDC2 PE = 1 SV = 2 Double-stranded RNA-bindingprotein Staufen homolog 63 kDa 0 15 4 4 1 OS = Homo sapiens OX = 9606 GN= STAU1 PE = 1 SV = 2 Dynactin subunit 2 OS = Homo sapiens GN = DCTN2 44kDa 0 6 0 2 PE = 1 SV = 4 Dynein heavy chain 10, axonemal OS = Homosapiens 515 kDa 0 1 0 2 OX = 9606 GN = DNAH10 PE = 1 SV = 4 EBNA1binding protein 2, isoform CRA_d OS = Homo 41 kDa 0 5 0 0 sapiens GN =EBNA1BP2 PE = 1 SV = 1 Ectonucleotide pyrophosphatase/phosphodiesterase105 kDa 0 0 0 2 family member 1 OS = Homo sapiens OX = 9606 GN = ENPP1PE = 1 SV = 2 EH domain-containing protein 4 OS = Homo sapiens 61 kDa 05 0 3 OX = 9606 GN = EHD4 PE = 1 SV = 1 Elongation factor Tu,mitochondrial OS = Homo sapiens 50 kDa 0 9 0 0 GN = TUFM PE = 1 SV = 2Emerin OS = Homo sapiens GN = EMD PE = 1 SV = 1 29 kDa 0 6 0 0 Epoxidehydrolase 1 OS = Homo sapiens OX = 9606 53 kDa 0 0 0 3 GN = EPHX1 PE = 1SV = 1 ER lumen protein-retaining receptor 3 OS = Homo sapiens 25 kDa 05 0 0 GN = KDELR3 PE = 2 SV = 1 Eukaryotic translation initiation factor3 subunit E 52 kDa 0 8 0 0 OS = Homo sapiens GN = EIF3E PE = 1 SV = 1Eukaryotic translation initiation factor 3 subunit F 38 kDa 0 14 0 0 OS= Homo sapiens GN = EIF3F PE = 1 SV = 1 Eukaryotic translationinitiation factor 3 subunit H 40 kDa 0 5 0 0 OS = Homo sapiens GN =EIF3H PE = 1 SV = 1 FACT complex subunit SPT16 OS = Homo sapiens 120 kDa0 7 0 0 GN = SUPT16H PE = 1 SV = 1 Far upstream element-binding protein1 OS = Homo 68 kDa 0 3 0 1 sapiens OX = 9606 GN = FUBP1 PE = 1 SV = 3Fascin OS = Homo sapiens GN = FSCN1 PE = 1 SV = 3 55 kDa 0 12 0 0Fibrous sheath-interacting protein 2 OS = Homo sapiens 781 kDa 0 0 2 1OX = 9606 GN = FSIP2 PE = 2 SV = 4 Filaggrin OS = Homo sapiens GN = FLGPE = 1 SV = 3 435 kDa 0 3 0 0 Filaggrin-2 OS = Homo sapiens GN = FLG2 PE= 1 SV = 1 248 kDa 0 2 0 0 Flotillin-2 OS = Homo sapiens GN = FLOT2 PE =1 SV = 1 53 kDa 0 13 0 0 Galectin-3 OS = Homo sapiens GN = LGALS3 PE = 1SV = 5 26 kDa 0 39 0 0 Galectin-3-binding protein OS = Homo sapiens OX =9606 65 kDa 0 4 0 3 GN = LGALS3BP PE = 1 SV = 1 Galectin-8 OS = Homosapiens OX = 9606 GN = LGALS8 36 kDa 0 5 2 0 PE = 1 SV = 4 Glutamatedehydrogenase 1, mitochondrial OS = Homo 61 kDa 0 4 0 0 sapiens OX =9606 GN = GLUD1 PE = 1 SV = 2 Glutathione peroxidase 1 OS = Homo sapiensGN = GPX1 22 kDa 0 3 0 2 PE = 1 SV = 4 GlycylpeptideN-tetradecanoyltransferase 2 OS = Homo 57 kDa 0 2 0 0 sapiens GN = NMT2PE = 1 SV = 1 Glypican-6 OS = Homo sapiens GN = GPC6 PE = 1 SV = 1 63kDa 0 14 0 0 Golgi-associated plant pathogenesis-related protein 1 17kDa 0 20 0 0 OS = Homo sapiens GN = GLIPR2 PE = 1 SV = 3 Gremlin-1 OS =Homo sapiens OX = 9606 GN = GREM1 21 kDa 0 0 3 0 PE = 1 SV = 1 Guaninenucleotide-binding protein G(I)/G(S)/G(O) 8 kDa 0 6 0 0 subunit gamma-12OS = Homo sapiens GN = GNG12 PE = 1 SV = 3 Guanine nucleotide-bindingprotein G(k) subunit alpha 41 kDa 0 17 0 0 OS = Homo sapiens GN = GNAI3PE = 1 SV = 3 Hamartin OS = Homo sapiens GN = TSC1 PE = 1 SV = 2 130 kDa0 0 0 2 Heat shock-related 70 kDa protein 2 OS = Homo sapiens 70 kDa 0 00 24 OX = 9606 GN = HSPA2 PE = 1 SV = 1 Hemicentin-2 OS = Homo sapiensOX = 9606 542 kDa 0 0 0 0 GN = HMCN2 PE = 2 SV = 3 Heparan sulfateglucosamine 3-O-sulfotransferase 6 37 kDa 0 3 0 0 OS = Homo sapiens GN =HS3ST6 PE = 1 SV = 2 Heterogeneous nuclear ribonucleoprotein A0 OS =Homo 31 kDa 0 15 0 0 sapiens GN = HNRNPA0 PE = 1 SV = 1 Heterogeneousnuclear ribonucleoprotein H2 OS = Homo 49 kDa 0 14 0 0 sapiens GN =HNRNPH2 PE = 1 SV = 1 Homeobox protein Hox-B3 OS = Homo sapiens OX =9606 44 kDa 0 5 0 0 GN = HOXB3 PE = 2 SV = 2 Hornerin OS = Homo sapiensGN = HRNR PE = 1 SV = 2 282 kDa 0 15 0 0 Hsc70-interacting protein(Fragment) OS = Homo sapiens 16 kDa 0 2 0 0 GN = ST13 PE = 1 SV = 1Hyaluronan and proteoglycan link protein 1 OS = Homo 40 kDa 0 0 0 0sapiens OX = 9606 GN = HAPLN1 PE = 2 SV = 2 Hyaluronan and proteoglycanlink protein 3 OS = Homo 41 kDa 0 6 0 0 sapiens GN = HAPLN3 PE = 2 SV =1 Intercellular adhesion molecule 1 OS = Homo sapiens 58 kDa 0 16 0 0 GN= ICAM1 PE = 1 SV = 2 Interleukin enhancer-binding factor 2 OS = Homosapiens 39 kDa 0 29 0 0 GN = ILF2 PE = 1 SV = 1 Isocitrate dehydrogenase[NADP] cytoplasmic 47 kDa 0 7 0 0 OS = Homo sapiens GN = IDH1 PE = 1 SV= 2 Isoform 1 of Gamma-adducin OS = Homo sapiens 76 kDa 0 7 0 1 OX =9606 GN = ADD3 Isoform 1 of Polypyrimidine tract-binding protein 3 57kDa 0 5 0 3 OS = Homo sapiens OX = 9606 GN = PTBP3 Isoform 1 of Synapticfunctional regulator FMR1 67 kDa 0 5 0 1 OS = Homo sapiens OX = 9606 GN= FMR1 Isoform 2 of 2,4-dienoyl-CoA reductase, mitochondrial 35 kDa 0 80 1 OS = Homo sapiens OX = 9606 GN = DECR1 Isoform 2 of A-kinase anchorprotein 13 OS = Homo 308 kDa 0 2 0 0 sapiens OX = 9606 GN = AKAP13Isoform 2 of Ankyrin repeat domain-containing protein 274 kDa 0 0 0 0 17OS = Homo sapiens OX = 9606 GN = ANKRD17 Isoform 2 of AP-2 complexsubunit mu OS = Homo 49 kDa 0 9 0 1 sapiens OX = 9606 GN = AP2M1 Isoform2 of Bcl-2-associated transcription factor 1 106 kDa 0 4 0 0 OS = Homosapiens OX = 9606 GN = BCLAF1 Isoform 2 of Cadherin-2 OS = Homo sapiensOX = 9606 97 kDa 0 7 0 2 GN = CDH2 Isoform 2 of Calponin-1 OS = Homosapiens OX = 9606 31 kDa 0 1 5 8 GN = CNN1 Isoform 2 of ChromodomainY-like protein OS = Homo 61 kDa 0 4 0 0 sapiens GN = CDYL Isoform 2 ofCollagen alpha-1(VII) chain OS = Homo 292 kDa 0 70 2 1 sapiens OX = 9606GN = COE7A1 Isoform 2 of Cyclin-Y OS = Homo sapiens GN = CCNY 37 kDa 0 30 0 Isoform 2 of Cysteine-rich protein 2 OS = Homo sapiens 30 kDa 0 5 00 OX = 9606 GN = CRIP2 Isoform 2 of DnaJ homolog subfamily C member 1086 kDa 0 4 0 0 OS = Homo sapiens GN = DNAJC10 Isoform 2 of Fibroblastgrowth factor 2 OS = Homo 23 kDa 0 13 3 1 sapiens OX = 9606 GN = FGF2Isoform 2 of Fibulin-2 OS = Homo sapiens OX = 9606 132 kDa 0 6 0 0 GN =FBLN2 Isoform 2 of GDNF family receptor alpha-1 OS = Homo 51 kDa 0 8 0 0sapiens GN = GFRA1 Isoform 2 of Glycogen phosphorylase, liver form 93kDa 0 2 0 1 OS = Homo sapiens OX = 9606 GN = PYGL Isoform 2 of Golginsubfamily A member 5 OS = Homo 78 kDa 0 0 0 0 sapiens OX = 9606 GN =GOLGA5 Isoform 2 of H/ACA ribonucleoprotein complex subunit 21 kDa 0 1 12 1 OS = Homo sapiens OX = 9606 GN = GAR1 Isoform 2 of Helicase SRCAP OS= Homo sapiens 337 kDa 0 2 0 0 OX = 9606 GN = SRCAP Isoform 2 ofHistone-binding protein RBBP4 OS = Homo 48 kDa 0 6 0 0 sapiens OX = 9606GN = RBBP4 Isoform 2 of Histone-lysine N-methyltransferase, H3 267 kDa 06 0 1 lysine-36 and H4 lysine-20 specific OS = Homo sapiens OX = 9606 GN= NSD1 Isoform 2 of Insulin-like growth factor-binding protein 7 29 kDa0 54 13 7 OS = Homo sapiens OX = 9606 GN = IGFBP7 Isoform 2 ofInterferon-inducible double-stranded RNA- 33 kDa 0 4 1 1 dependentprotein kinase activator A OS = Homo sapiens OX = 9606 GN = PRKRAIsoform 2 of KH domain-containing, RNA-binding, 30 kDa 0 8 0 0 signaltransduction-associated protein 3 OS = Homo sapiens GN = KHDRBS3 Isoform2 of Lactadherin OS = Homo sapiens 35 kDa 0 25 2 1 GN = MFGE8 Isoform 2of L-amino-acid oxidase OS = Homo sapiens 65 kDa 0 2 0 1 OX = 9606 GN =IL4I1 Isoform 2 of Macrophage-capping protein OS = Homo 37 kDa 0 3 0 0sapiens GN = CAPG Isoform 2 of Matrilin-2 OS = Homo sapiens OX = 9606105 kDa 0 37 8 5 GN = MATN2 Isoform 2 of Mesoderm-specific transcripthomolog 38 kDa 0 0 0 4 protein OS = Homo sapiens OX = 9606 GN = MESTIsoform 2 of Microtubule-associated protein 1A 306 kDa 0 4 2 3 OS = Homosapiens OX = 9606 GN = MAP1A Isoform 2 of Midkine OS = Homo sapiens OX =9606 10 kDa 0 4 2 2 GN = MDK Isoform 2 of Monoacylglycerol lipase ABHD1246 kDa 0 2 0 0 OS = Homo sapiens GN = ABHD12 Isoform 2 of Multidrugresistance protein 1 OS = Homo 134 kDa 0 1 0 2 sapiens OX = 9606 GN =ABCB1 Isoform 2 of Myosin-11 OS = Homo sapiens GN = MYH11 228 kDa 0 16137 0 Isoform 2 of Myosin-14 OS = Homo sapiens OX = 9606 232 kDa 0 141 2626 GN = MYH14 Isoform 2 of NADH dehydrogenase [ubiquinone] 1 beta 14 kDa0 1 0 2 subcomplex subunit 4 OS = Homo sapiens OX = 9606 GN = NDUFB4Isoform 2 of Nuclear receptor corepressor 1 OS = Homo 259 kDa 0 0 2 0sapiens OX = 9606 GN = NCOR1 Isoform 2 of Periostin OS = Homo sapiens OX= 9606 87 kDa 0 0 5 5 GN = POSTN Isoform 2 of Pescadillo homolog OS =Homo sapiens 67 kDa 0 5 1 0 OX = 9606 GN = PES1 Isoform 2 ofPlatelet-derived growth factor subunit B 26 kDa 0 1 3 1 OS = Homosapiens OX = 9606 GN = PDGFB Isoform 2 of Protein ELYS OS = Homo sapiensOX = 9606 256 kDa 0 0 2 0 GN = AHCTF1 Isoform 2 of Ran-binding protein 3OS = Homo sapiens 60 kDa 0 5 0 0 OX = 9606 GN = RANBP3 Isoform 2 ofRegulator of chromosome condensation 48 kDa 0 16 0 0 OS = Homo sapiensOX = 9606 GN = RCC1 Isoform 2 of Retinol-binding protein 1 OS = Homo 17kDa 0 0 0 2 sapiens OX = 9606 GN = RBP1 Isoform 2 of RNA-binding protein39 OS = Homo sapiens 59 kDa 0 0 0 2 GN = RBM39 Isoform 2 of RNA-bindingprotein 8A OS = Homo sapiens 20 kDa 0 5 0 1 OX = 9606 GN = RBM8A Isoform2 of Ryanodine receptor 1 OS = Homo sapiens 565 kDa 0 1 0 2 OX = 9606 GN= RYR1 Isoform 2 of SCO-spondin OS = Homo sapiens OX = 9606 139 kDa 0 30 0 GN = SSPO Isoform 2 of Semaphorin-7A OS = Homo sapiens 73 kDa 0 3 13 OX = 9606 GN = SEMA7A Isoform 2 of Septin-8 OS = Homo sapiens OX =9606 50 kDa 0 12 0 3 GN = SEPT8 Isoform 2 of Serine/threonine-proteinphosphatase 28 kDa 0 5 0 1 PGAM5, mitochondrial OS = Homo sapiens OX =9606 GN = PGAM5 Isoform 2 of SH3 domain-containing kinase-binding 69 kDa0 2 0 2 protein 1 OS = Homo sapiens OX = 9606 GN = SH3KBP1 Isoform 2 ofSignal recognition particle receptor subunit 67 kDa 0 3 0 1 alpha OS =Homo sapiens OX = 9606 GN = SRPRA Isoform 2 of Signal-inducedproliferation-associated 1- 197 kDa 0 0 0 0 like protein 1 OS = Homosapiens OX = 9606 GN = SIPA1L1 Isoform 2 of Sorting nexin-27 OS = Homosapiens 60 kDa 0 4 0 0 GN = SNX27 Isoform 2 of Spectrin beta chain,erythrocytic OS = Homo 268 kDa 0 8 0 2 sapiens OX = 9606 GN = SPTBIsoform 2 of Testis-expressed protein 10 OS = Homo 104 kDa 0 2 0 0sapiens OX = 9606 GN = TEX10 Isoform 2 of Tissue factor pathwayinhibitor 2 OS = Homo 26 kDa 0 9 6 0 sapiens OX = 9606 GN = TFPI2Isoform 2 of Tyrosine-protein kinase BAZ1B OS = Homo 170 kDa 0 4 0 0sapiens OX = 9606 GN = BAZ1B Isoform 2 of UDP-glucuronosyltransferase1-6 30 kDa 0 4 0 0 OS = Homo sapiens OX = 9606 GN = UGT1A6 Isoform 2 ofVesicle-associated membrane protein- 33 kDa 0 3 0 1 associated protein AOS = Homo sapiens OX = 9606 GN = VAPA Isoform 2 of Voltage-dependentcalcium channel subunit 123 kDa 0 2 1 0 alpha-2/delta-l OS = Homosapiens OX = 9606 GN = CACNA2D1 Isoform 2 of Y-box-binding protein 3 OS= Homo sapiens 32 kDa 0 15 0 1 OX = 9606 GN = YBX3 Isoform 2 of Zincfinger homeobox protein 4 OS = Homo 397 kDa 0 2 0 0 sapiens OX = 9606 GN= ZFHX4 Isoform 3 of 1-phosphatidylinositol 4,5-bisphosphate 136 kDa 0 30 0 phosphodiesterase beta-4 OS = Homo sapiens OX = 9606 GN = PLCB4Isoform 3 of Alpha-adducin OS = Homo sapiens 84 kDa 0 2 0 0 GN = ADD1Isoform 3 of Cytoskeleton-associated protein 5 226 kDa 0 3 0 0 OS = Homosapiens OX = 9606 GN = CKAP5 Isoform 3 of DnaJ homolog subfamily Cmember 11 57 kDa 0 2 0 1 OS = Homo sapiens GN = DNAJC11 Isoform 3 of E3ubiquitin-protein ligase CHFR 69 kDa 0 0 0 0 OS = Homo sapiens OX = 9606GN = CHFR Isoform 3 of H/ACA ribonucleoprotein complex subunit 48 kDa 013 1 1 DKC1 OS = Homo sapiens OX = 9606 GN = DKC1 Isoform 3 ofHistone-lysine N-methyltransferase 2D 594 kDa 0 1 0 1 OS = Homo sapiensOX = 9606 GN = KMT2D Isoform 3 of Latent-transforming growth factorbeta- 169 kDa 0 34 0 0 binding protein 4 OS = Homo sapiens OX = 9606 GN= LTBP4 Isoform 3 of Malate dehydrogenase, cytoplasmic 39 kDa 0 3 0 0 OS= Homo sapiens OX = 9606 GN = MDH1 Isoform 3 of Putative oxidoreductaseGLYR1 OS = Homo 60 kDa 0 4 0 0 sapiens OX = 9606 GN = GLYR1 Isoform 3 ofScaffold attachment factor B1 OS = Homo 103 kDa 0 3 0 0 sapiens GN =SAFB Isoform 3 of Torsin-1A-interacting protein 1 OS = Homo 66 kDa 0 5 00 sapiens GN = TOR1AIP1 Isoform 4 of CD109 antigen OS = Homo sapiens 160kDa 0 18 0 0 GN = CD109 Isoform 4 of FYVE and coiled-coildomain-containing 169 kDa 0 0 0 0 protein 1 OS = Homo sapiens OX = 9606GN = FYCO1 Isoform 4 of IQ domain-containing protein N OS = Homo 147 kDa0 0 0 3 sapiens OX = 9606 GN = IQCN Isoform 4 of Kinesin-like proteinKIF24 OS = Homo 129 kDa 0 2 0 0 sapiens OX = 9606 GN = KIF24 Isoform 4of Latent-transforming growth factor beta- 187 kDa 0 1 4 1 bindingprotein 1 OS = Homo sapiens OX = 9606 GN = LTBP1 Isoform 4 of Proteindiaphanous homolog 3 OS = Homo 136 kDa 0 2 0 0 sapiens GN = DIAPH3Isoform 5 of E1A-binding protein p400 OS = Homo 340 kDa 0 1 0 0 sapiensOX = 9606 GN = EP400 Isoform 5 of Immunoglobulin-like and fibronectintype 384 kDa 0 3 0 2 III domain-containing protein 1 OS = Homo sapiensOX = 9606 GN = IGFN1 Isoform 5 of LIM domain only protein 7 OS = Homo158 kDa 0 8 0 0 sapiens GN = LMO7 Isoform 5 of Papilin OS = Homo sapiensOX = 9606 136 kDa 0 17 1 1 GN = PAPLN Isoform 6 of Treacle protein OS =Homo sapiens 148 kDa 0 2 0 0 OX = 9606 GN = TCOF1 Isoform B of Collagenalpha-1(XI) chain OS = Homo 182 kDa 0 0 1 2 sapiens OX = 9606 GN =COL11Al Isoform B of Collagen alpha-6(IV) chain OS = Homo 164 kDa 0 5 20 sapiens OX = 9606 GN = COL4A6 Isoform B of DnaJ homolog subfamily Bmember 6 27 kDa 0 0 1 2 OS = Homo sapiens OX = 9606 GN = DNAJB6 IsoformB of Methyl-CpG-binding protein 2 OS = Homo 53 kDa 0 11 1 0 sapiens OX =9606 GN = MECP2 Isoform B of Ras-related C3 botulinum toxin substrate 123 kDa 0 3 0 0 OS = Homo sapiens OX = 9606 GN = RAC1 Isoform B ofTransforming growth factor beta-2 51 kDa 0 5 1 2 proprotein OS = Homosapiens OX = 9606 GN = TGFB2 Isoform Beta-3B of Integrin beta-3 OS =Homo sapiens 86 kDa 0 4 0 2 OX = 9606 GN = ITGB3 Isoform C of Fibulin-1OS = Homo sapiens OX = 9606 74 kDa 0 13 0 8 GN = FBLN1 Isoform Long ofProteasome subunit alpha type-1 30 kDa 0 2 0 2 OS = Homo sapiens OX =9606 GN = PSMA1 Isoform Non-brain of Clathrin light chain A OS = Homo 24kDa 0 17 0 2 sapiens OX = 9606 GN = CLTA Isoform Short of Lamininsubunit gamma-2 OS = Homo 122 kDa 0 5 0 1 sapiens OX = 9606 GN = LAMC2Keratin, type I cytoskeletal 14 OS = Homo sapiens 52 kDa 0 94 0 18 OX =9606 GN = KRT14 PE = 1 SV = 4 Keratin, type I cytoskeletal 16 OS = Homosapiens 51 kDa 0 99 0 0 GN = KRT16 PE = 1 SV = 4 Keratin, type Icytoskeletal 19 OS = Homo sapiens 44 kDa 0 753 0 0 GN = KRT19 PE = 1 SV= 4 Keratin, type II cuticular Hb5 OS = Homo sapiens 56 kDa 0 3 0 0 GN =KRT85 PE = 1 SV = 1 Keratin, type II cytoskeletal 1 OS = Homo sapiens 66kDa 0 497 0 0 GN = KRT1 PE = 1 SV = 6 Keratin, type II cytoskeletal 5 OS= Homo sapiens 62 kDa 0 93 0 0 GN = KRT5 PE = 1 SV = 3 Keratin, type IIcytoskeletal 6B OS = Homo sapiens 60 kDa 0 104 18 0 OX = 9606 GN = KRT6BPE = 1 SV = 5 Keratin, type II cytoskeletal 7 OS = Homo sapiens 51 kDa 00 0 135 OX = 9606 GN = KRT7 PE = 1 SV = 5 Kinesin-like protein KIFC2 OS= Homo sapiens OX = 9606 90 kDa 0 2 0 1 GN = KIFC2 PE = 2 SV = 1Ladinin-1 OS = Homo sapiens OX = 9606 GN = LAD1 57 kDa 0 0 0 6 PE = 1 SV= 2 Laminin subunit beta-3 OS = Homo sapiens GN = LAMB3 130 kDa 0 10 0 0PE = 1 SV = 1 Latent-transforming growth factor beta-binding protein 2190 kDa 0 23 0 0 OS = Homo sapiens GN = LTBP2 PE = 1 SV = 1 Leucinezipper protein 1 OS = Homo sapiens OX = 9606 120 kDa 0 5 0 0 GN = LUZP1PE = 1 SV = 2 Lipoamide acyltransferase component of branched-chain 53kDa 0 6 4 8 alpha-keto acid dehydrogenase complex, mitochondrial OS =Homo sapiens OX = 9606 GN = DBT PE = 1 SV = 3 Low-density lipoproteinreceptor-related protein 1B 515 kDa 0 2 0 0 OS = Homo sapiens GN = LRP1BPE = 1 SV = 2 Low-density lipoprotein receptor-related protein 2 522 kDa0 0 0 2 OS = Homo sapiens OX = 9606 GN = LRP2 PE = 1 SV = 3 Lysyloxidase homolog 1 OS = Homo sapiens OX = 9606 63 kDa 0 24 1 0 GN = LOXL1PE = 1 SV = 2 Magnesium transporter protein 1 OS = Homo sapiens 42 kDa 04 1 1 GN = MAGT1 PE = 1 SV = 1 MARCKS-related protein OS = Homo sapiens20 kDa 0 4 0 0 GN = MARCKSL1 PE = 1 SV = 2 Metastasis-associated proteinMTA2 OS = Homo sapiens 75 kDa 0 5 0 0 GN = MTA2 PE = 1 SV = 1 Microsomalglutathione S-transferase 1 OS = Homo 18 kDa 0 0 0 3 sapiens OX = 9606GN = MGST1 PE = 1 SV = 1 Mitochondrial GTPase 1 OS = Homo sapiens OX =9606 37 kDa 0 0 0 2 PE = 3 SV = 1 MKI67 FHA domain-interacting nucleolar20 kDa 0 6 0 0 phosphoprotein (Fragment) OS = Homo sapiens GN = NIFK PE= 1 SV = 1 Mucin-16 OS = Homo sapiens OX = 9606 GN = MUC16 1519 kDa 0 02 4 PE = 1 SV = 3 Myelin expression factor 2 OS = Homo sapiens OX = 960664 kDa 0 2 0 0 GN = MYEF2 PE = 1 SV = 3 Myosin light chain 6B OS = Homosapiens OX = 9606 23 kDa 0 0 6 0 GN = MYL6B PE = 1 SV = 1 Myristoylatedalanine-rich C-kinase substrate OS = Homo 32 kDa 0 11 0 0 sapiens GN =MARCKS PE = 1 SV = 4 N-acylneuraminate cytidylyltransferase OS = Homo 48kDa 0 6 0 0 sapiens GN = CMAS PE = 1 SV = 2 NAD(P) transhydrogenase,mitochondrial OS = Homo 100 kDa 0 3 0 0 sapiens GN = NNT PE = 1 SV = 1Nestin OS = Homo sapiens GN = NES PE = 1 SV = 2 177 kDa 0 18 0 0Neurabin-2 OS = Homo sapiens GN = PPP1R9BPE = 1 89 kDa 0 3 0 0 SV = 1Neurobeachin OS = Homo sapiens OX = 9606GN = NBEA 328 kDa 0 0 0 3 PE = 1SV = 3 Nicotinate-nucleotide pyrophosphorylase [carboxylating] 31 kDa 00 0 4 OS = Homo sapiens OX = 9606 GN = QPRT PE = 1 SV = 3 Non-syndromichearing impairment protein 5 OS = Homo 55 kDa 0 2 0 0 sapiens GN = DFNA5PE = 1 SV = 2 Nuclear receptor-binding protein OS = Homo sapiens 61 kDa0 5 0 0 GN = NRBP1 PE = 1 SV = 1 Nucleolar complex protein 3 homolog OS= Homo sapiens 93 kDa 0 5 0 0 GN = NOC3L PE = 1 SV = 1 Nucleolar complexprotein 4 homolog OS = Homo sapiens 58 kDa 0 5 0 0 GN = NOC4L PE = 1 SV= 1 Nucleolar protein 58 OS = Homo sapiens GN = NOP58 60 kDa 0 6 0 0 PE= 1 SV = 1 Nucleolar transcription factor 1 OS = Homo sapiens 87 kDa 0 20 0 GN = UBTF PE = 1 SV = 1 Nucleoplasmin-3 OS = Homo sapiens GN = NPM3PE = 1 19 kDa 0 3 0 0 SV = 3 Palladin OS = Homo sapiens OX = 9606 GN =PALLD 151 kDa 0 7 0 6 PE = 1 SV = 3 PDZ and LIM domain protein 1 OS =Homo sapiens 36 kDa 0 0 0 4 OX = 9606 GN = PDLIM1 PE = 1 SV = 4 PDZ andLIM domain protein 4 OS = Homo sapiens 35 kDa 0 3 0 0 GN = PDLIM4 PE = 1SV = 2 Pentraxin-related protein PTX3 OS = Homo sapiens 42 kDa 0 23 0 0GN = PTX3 PE = 1 SV = 3 Peptidyl-prolyl cis-trans isomerase B OS = Homosapiens 24 kDa 0 52 0 0 GN = PPIB PE = 1 SV = 2 Peptidyl-prolylcis-trans isomerase FKBP10 OS = Homo 64 kDa 0 3 0 0 sapiens GN = FKBP10PE = 1 SV = 1 Peptidyl-prolyl cis-trans isomerase FKBP3 OS = Homo 25 kDa0 8 0 0 sapiens GN = FKBP3 PE = 1 SV = 1 Periaxin OS = Homo sapiens OX =9606 GN = PRX PE = 1 155 kDa 0 0 0 2 SV = 2 Periodic tryptophan protein1 homolog OS = Homo 56 kDa 0 2 0 0 sapiens OX = 9606 GN = PWP1 PE = 1 SV= 1 Peroxiredoxin-1 (Fragment) OS = Homo sapiens 19 kDa 0 13 0 0 GN =PRDX1 PE = 1 SV = 1 Phosphoglycerate mutase 1 OS = Homo sapiens 29 kDa 011 0 0 GN = PGAM1 PE = 1 SV = 2 Pinin OS = Homo sapiens OX = 9606 GN =PNN PE = 1 82 kDa 0 6 2 4 SV = 5 Platelet-activating factoracetylhydrolase IB subunit 47 kDa 0 4 0 0 alpha OS = Homo sapiens OX =9606 GN = PAFAH1B1 PE = 1 SV = 2 Poly [ADP-ribose] polymerase 1 OS =Homo sapiens 113 kDa 0 30 0 0 GN = PARP1 PE = 1 SV = 4Poly(U)-binding-splicing factor PUF60 (Fragment) 57 kDa 0 3 0 0 OS =Homo sapiens GN = PUF60 PE = 1 SV = 1 Polymerase delta-interactingprotein 3 OS = Homo sapiens 48 kDa 0 6 0 0 GN = POLDIP3 PE = 1 SV = 1Polymerase I and transcript release factor OS = Homo 43 kDa 0 89 0 0sapiens GN = PTRF PE = 1 SV = 1 Polyubiquitin-B OS = Homo sapiens GN =UBBPE = 1 17 kDa 0 111 0 0 SV = 1 POU domain, class 3, transcriptionfactor 3 OS = Homo 50 kDa 0 20 0 0 sapiens GN = POU3F3 PE = 2 SV = 2 PRdomain zinc finger protein 8 OS = Homo sapiens 72 kDa 0 2 0 0 GN = PRDM8PE = 1 SV = 3 Prefoldin subunit 6 OS = Homo sapiens GN = PFDN6 15 kDa 03 0 0 PE = 1 SV = 1 Probable ATP-dependent RNA helicase DDX27 87 kDa 0 40 0 OS = Homo sapiens GN = DDX27 PE = 1 SV = 1 Probable globaltranscription activator SNF2L1 123 kDa 0 6 0 0 OS = Homo sapiens OX =9606 GN = SMARCA1 PE = 1 SV = 2 Probable maltase-glucoamylase 2 OS =Homo sapiens 278 kDa 0 0 0 2 OX = 9606 GN = MGAM2 PE = 2 SV = 3Procollagen galactosyltransferase 1 OS = Homo sapiens 72 kDa 0 3 0 0 GN= COLGALT1 PE = 1 SV = 1 Procollagen-lysine,2-oxoglutarate 5-dioxygenase3 85 kDa 0 3 0 0 OS = Homo sapiens GN = PLOD3 PE = 1 SV = 1Prolow-density lipoprotein receptor-related protein 1 505 kDa 0 2 0 0 OS= Homo sapiens GN = LRP1 PE = 1 SV = 2 Protein disulfide-isomerase OS =Homo sapiens 53 kDa 0 30 0 0 GN = P4HB PE = 1 SV = 2 Protein GREB1 OS =Homo sapiens GN = GREB1PE = 2 216 kDa 0 0 0 2 SV = 1 Protein kinase Cdelta-binding protein OS = Homo sapiens 31 kDa 0 20 0 0 GN = PRKCDBP PE= 1 SV = 1 Protein MAK16 homolog OS = Homo sapiens 35 kDa 0 2 0 0 GN =MAK16 PE = 1 SV = 2 Protein S100-A10 OS = Homo sapiens GN = S100A10 11kDa 0 11 0 0 PE = 1 SV = 2 Protein S100-A13 OS = Homo sapiens GN =S100A13 11 kDa 0 5 0 0 PE = 1 SV = 1 Protein S100-A9 OS = Homo sapiensGN = S100A9 PE = 1 13 kDa 0 12 0 0 SV = 1 Protein-glutaminegamma-glutamyltransferase E 77 kDa 0 2 0 0 OS = Homo sapiens GN = TGM3PE = 1 SV = 4 Pumilio homolog 3 OS = Homo sapiens GN = PUM3 PE = 1 74kDa 0 6 0 0 SV = 3 Raftlin OS = Homo sapiens GN = RFTN1 PE = 1 SV = 4 63kDa 0 3 0 0 Ras GTPase-activating-like protein IQGAP1 OS = Homo 189 kDa0 135 0 0 sapiens GN = IQGAP1 PE = 1 SV = 1 Ras-related protein Rab-10OS = Homo sapiens 23 kDa 0 13 0 0 GN = RAB10 PE = 1 SV = 1 Ras-relatedprotein Rab-14 (Fragment) OS = Homo 20 kDa 0 8 0 0 sapiens GN = RAB14 PE= 1 SV = 1 Ras-related protein Rab-2A OS = Homo sapiens 24 kDa 0 5 0 1GN = RAB2A PE = 1 SV = 1 Ras-related protein Ral-A OS = Homo sapiens GN= RALA 24 kDa 0 2 0 0 PE = 1 SV = 1 Regulation of nuclear pre-mRNAdomain-containing 37 kDa 0 2 0 0 protein 1B OS = Homo sapiens GN =RPRD1B PE = 1 SV = 1 Replication protein A 32 kDa subunit OS = Homosapiens 29 kDa 0 3 0 1 GN = RPA2 PE = 1 SV = 1 Replication protein A 70kDa DNA-binding subunit 68 kDa 0 4 0 0 OS = Homo sapiens GN = RPA1 PE =1 SV = 2 Reticulocalbin-1 OS = Homo sapiens GN = RCN1 PE = 1 39 kDa 0 70 0 SV = 1 Retinoic acid-induced protein 3 OS = Homo sapiens 40 kDa 0 00 2 OX = 9606 GN = GPRC5A PE = 1 SV = 2 Rho GTPase-activating protein 1OS = Homo sapiens 50 kDa 0 8 0 0 GN = ARHGAP1 PE = 1 SV = 1 Ribosomalprotein L19 OS = Homo sapiens GN = RPL19 23 kDa 0 21 0 0 PE = 1 SV = 1Ribosome biogenesis protein BRX1 homolog OS = Homo 41 kDa 0 5 0 3sapiens OX = 9606 GN = BRIX1 PE = 1 SV = 2 Ribosome biogenesis proteinWDR12 OS = Homo sapiens 48 kDa 0 3 0 0 GN = WDR12 PE = 1 SV = 2 Ribosomebiogenesis regulatory protein homolog 41 kDa 0 11 0 0 OS = Homo sapiensGN = RRS1 PE = 1 SV = 2 Ribosome production factor 2 homolog OS = Homo36 kDa 0 3 0 0 sapiens GN = RPF2 PE = 1 SV = 2 RNA-binding protein 28 OS= Homo sapiens OX = 9606 86 kDa 0 2 0 2 GN = RBM28 PE = 1 SV = 3RNA-binding protein 3 OS = Homo sapiens GN = RBM3 17 kDa 0 3 0 0 PE = 1SV = 1 rRNA 2′-O-methyltransferase fibrillarin OS = Homo 34 kDa 0 23 0 0sapiens GN = FBL PE = 1 SV = 2 Selenoprotein H OS = Homo sapiens GN =C11orf31 PE = 1 13 kDa 0 2 0 0 SV = 1 Semaphorin-3C OS = Homo sapiens GN= SEMA3C PE = 2 85 kDa 0 3 0 0 SV = 2 Serine protease 23 OS = Homosapiens OX = 9606 43 kDa 0 7 7 5 GN = PRSS23 PE = 1 SV = 1 Serineprotease HTRA3 OS = Homo sapiens OX = 9606 49 kDa 0 3 3 0 GN = HTRA3 PE= 1 SV = 2 Serine/threonine-protein phosphatase 2A catalytic 36 kDa 0 40 0 subunit beta isoform OS = Homo sapiens GN = PPP2CB PE = 1 SV = 1Serine/threonine-protein phosphatase PP1-beta catalytic 37 kDa 0 22 0 0subunit OS = Homo sapiens GN = PPP1CB PE = 1 SV = 3 Serpin B3 OS = Homosapiens GN = SERPINB3PE = 1 45 kDa 0 4 0 0 SV = 2 SH3 domain-bindingprotein 1 OS = Homo sapiens 76 kDa 0 2 0 0 GN = SH3BP1 PE = 1 SV = 3Signal peptidase complex subunit 1 OS = Homo sapiens 12 kDa 0 2 0 0 GN =SPCS1 PE = 1 SV = 4 Signal peptidase complex subunit 3 OS = Homo sapiens20 kDa 0 5 0 0 GN = SPCS3 PE = 1 SV = 1 Signal recognition particle 14kDa protein OS = Homo 15 kDa 0 9 0 0 sapiens GN = SRP14 PE = 1 SV = 2Signal-induced proliferation-associated 1-like protein 2 190 kDa 0 0 0 2OS = Homo sapiens OX = 9606 GN = SIPA1L2 PE = 1 SV = 2 Single-strandedDNA-binding protein, mitochondrial 17 kDa 0 24 0 0 OS = Homo sapiens GN= SSBP1 PE = 1 SV = 1 SNW domain-containing protein 1 OS = Homo sapiens61 kDa 0 4 0 0 GN = SNW1 PE = 1 SV = 1 Solute carrier family 2,facilitated glucose transporter 54 kDa 0 9 0 0 member 1 OS = Homosapiens GN = SLC2A1 PE = 1 SV = 2 SPARC (Fragment) OS = Homo sapiens GN= SPARC 17 kDa 0 2 0 0 PE = 1 SV = 1 Spermatogenesis-associatedserine-rich protein 2 60 kDa 0 2 0 0 OS = Homo sapiens GN = SPATS2 PE =1 SV = 1 Sphingosine-1-phosphate lyase 1 OS = Homo sapiens 64 kDa 0 2 00 GN = SGPL1 PE = 1 SV = 3 Splicing factor 3B subunit 2 OS = Homosapiens 98 kDa 0 7 0 0 GN = SF3B2 PE = 1 SV = 1 SRAstem-loop-interacting RNA-binding protein, 14 kDa 0 2 0 0 mitochondrialOS = Homo sapiens GN = SLIRP PE = 1 SV = 1 Structural maintenance ofchromosomes protein 3 142 kDa 0 11 0 0 OS = Homo sapiens GN = SMC3 PE =1 SV = 2 SWI/SNF-related matrix-associated actin-dependent 122 kDa 0 170 0 regulator of chromatin subfamily A member 5 OS = Homo sapiens GN =SMARCA5 PE = 1 SV = 1 SWI/SNF-related matrix-associated actin-dependent47 kDa 0 4 0 0 regulator of chromatin subfamily E member 1 OS = Homosapiens OX = 9606 GN = SMARCE1 PE = 1 SV = 2 SWISS-PROT: P01044-1 (Bostaurus) Isoform HMW of 69 kDa 0 3 0 0 Kininogen-1 precursor SWISS-PROT:P02768-1 Tax_Id = 9606 69 kDa 0 0 5 13 Gene_Symbol = ALB Isoform 1 ofSerum albumin precursor SWISS-PROT: Q3SZR3 (Bos taurus) Alpha-1-acid 23kDa 0 2 0 0 glycoprotein precursor SWISS-PROT: Q3TTY5 Tax_Id = 10090 71kDa 0 27 7 7 Gene_Symbol = Krt2 Keratin, type II cytoskeletal 2epidermal SWISS-PROT: Q9D312 Tax_Id = 10090 49 kDa 0 6 0 4 Gene_Symbol =Krt20 Keratin, type I cytoskeletal 20 SWISS-PROT: Q9QWL7 Tax_Id = 1009048 kDa 0 54 0 16 Gene_Symbol = Krt17 Keratin, type I cytoskeletal 17Synaptosomal-associated protein 23 OS = Homo sapiens 23 kDa 0 6 0 0 GN =SNAP23 PE = 1 SV = 1 Tau-tubulin kinase 2 OS = Homo sapiens OX = 9606182 kDa 0 0 0 0 GN = TTBK2 PE = 1 SV = 1 TBC1 domain family member 1(Fragment) OS = Homo 98 kDa 0 2 0 0 sapiens GN = TBC1D1 PE = 1 SV = 3T-complex protein 1 subunit alpha OS = Homo sapiens 60 kDa 0 98 0 0 GN =TCP1 PE = 1 SV = 1 Thyroid hormone receptor-associated protein 3 109 kDa0 11 0 0 OS = Homo sapiens GN = THRAP3 PE = 1 SV = 2 Tight junctionprotein ZO-1 OS = Homo sapiens 188 kDa 0 3 0 0 GN = TJP1 PE = 1 SV = 1Tight junction protein ZO-2 OS = Homo sapiens 141 kDa 0 3 0 0 GN = TJP2PE = 4 SV = 1 Transcription factor A, mitochondrial OS = Homo sapiens 29kDa 0 14 1 1 OX = 9606 GN = TFAM PE = 1 SV = 1 Transcription factorSOX-3 OS = Homo sapiens 45 kDa 0 2 0 0 GN = SOX3 PE = 1 SV = 2Transforming growth factor-beta-induced protein ig-h3 75 kDa 0 670 73 58OS = Homo sapiens OX = 9606 GN = TGFBI PE = 1 SV = 1 Translocatorprotein OS = Homo sapiens OX = 9606 19 kDa 0 0 0 3 GN = TSPO PE = 1 SV =3 Transmembrane protein 165 OS = Homo sapiens 35 kDa 0 5 0 1 GN =TMEM165 PE = 1 SV = 1 TREMBL: Q0V8M9; Q9TRI0 (Bos taurus) similar to 100kDa 0 7 0 0 inter-alpha (globulin) inhibitor H3 isoform 2 TREMBL:Q2KJC7; Q8HZM3 (Bos taurus) Periostin, 87 kDa 0 1 12 5 osteoblastspecific factor TREMBL: Q3SZH5 (Bos taurus) Similar to 45 kDa 0 11 4 2Angiotensinogen TREMBL: Q3T052; Q5EA67 (Bos taurus) Inter-alpha 102 kDa0 10 5 1 (Globulin) inhibitor H4 TREMBL: Q6ISB0 Keratin, hair, basic,4 - Homo sapiens 65 kDa 0 16 8 8 (Human). TREMBL: Q6NXH9 Tax_Id = 1009059 kDa 0 114 7 10 Gene_Symbol = Krt73 Keratin 73 TRIO andF-actin-binding protein OS = Homo sapiens 261 kDa 0 2 0 0 OX = 9606 GN =TRIOBP PE = 1 SV = 3 Tropomodulin-3 OS = Homo sapiens OX = 9606 40 kDa 047 21 13 GN = TMOD3 PE = 1 SV = 1 Tropomyosin 1 (Alpha), isoform CRA_fOS = Homo 37 kDa 0 162 0 0 sapiens GN = TPM1 PE = 1 SV = 1 Tropomyosin 1(Alpha), isoform CRA_m OS = Homo 29 kDa 0 118 0 0 sapiens GN = TPM1 PE =1 SV = 1 Tropomyosin alpha-3 chain OS = Homo sapiens 33 kDa 0 112 0 0 GN= TPM3 PE = 1 SV = 1 Tubby-related protein 2 (Fragment) OS = Homosapiens 24 kDa 0 0 0 2 OX = 9606 GN = TULP2 PE = 4 SV = 1 Twinfilin-1 OS= Homo sapiens OX = 9606 GN = TWF1 40 kDa 0 1 1 4 PE = 1 SV = 3Tyrosine-protein kinase OS = Homo sapiens GN = YES1 61 kDa 0 7 0 0 PE =1 SV = 1 Tyrosine--tRNA ligase OS = Homo sapiens GN = YARS 44 kDa 0 6 00 PE = 1 SV = 1 U1 small nuclear ribonucleoprotein A (Fragment) 28 kDa 07 0 0 OS = Homo sapiens GN = SNRPA PE = 1 SV = 1 U3 small nucleolarribonucleoprotein protein MPP10 79 kDa 0 6 0 0 OS = Homo sapiens GN =MPHOSPH10 PE = 1 SV = 2 U3 small nucleolar RNA-associated protein 14Homolog 88 kDa 0 2 0 0 A OS = Homo sapiens OX = 9606 GN = UTP14A PE = 1SV = 1 U4/U6.U5 tri-snRNP-associated protein 1 OS = Homo 90 kDa 0 4 0 0sapiens GN = SART1 PE = 1 SV = 1 UAP56-interacting factor OS = Homosapiens OX = 9606 36 kDa 0 3 0 0 GN = FYTTD1 PE = 1 SV = 3 Ubiquitincarboxyl-terminal hydrolase 24 OS = Homo 294 kDa 0 0 0 2 sapiens OX =9606 GN = USP24 PE = 1 SV = 3 Unconventional myosin-Id OS = Homo sapiensOX = 9606 116 kDa 0 0 2 0 GN = MYO1D PE = 1 SV = 2 Unconventionalmyosin-VI OS = Homo sapiens 145 kDa 0 26 0 0 GN = MYO6 PE = 1 SV = 1Unconventional myosin-XV OS = Homo sapiens 395 kDa 0 0 2 0 OX = 9606 GN= MYO15A PE = 1 SV = 2 Urokinase-type plasminogen activator OS = Homosapiens 47 kDa 0 8 0 0 GN = PLAU PE = 1 SV = 1 UV excision repairprotein RAD23 homolog B 43 kDa 0 5 0 0 OS = Homo sapiens GN = RAD23B PE= 1 SV = 1 Vacuolar protein sorting-associated protein 26A 38 kDa 0 5 03 OS = Homo sapiens OX = 9606 GN = VPS26A PE = 1 SV = 2 Very-long-chainenoyl-CoA reductase OS = Homo sapiens 36 kDa 0 6 0 0 GN = TECR PE = 1 SV= 1 Vesicle-trafficking protein SEC22b OS = Homo sapiens 25 kDa 0 6 0 0GN = SEC22B PE = 1 SV = 4 V-type proton ATPase subunit B, brain isoform57 kDa 0 16 0 0 OS = Homo sapiens GN = ATP6V1B2 PE = 1 SV = 3 V-typeproton ATPase subunit d 1 OS = Homo sapiens 45 kDa 0 22 0 0 GN =ATP6V0D1 PE = 1 SV = 1 Zinc finger protein 469 OS = Homo sapiens OX =9606 410 kDa 0 0 0 4 GN = ZNF469 PE = 2 SV = 3

Example 3—Proliferation of iPSCs on Amniotic Fluid Cell-Derived ECM

Induced pluripotent stem cells (iPSCs) were allowed to proliferate on anamniotic fluid cell-derived ECM (Matrix B) from Example 1 in cultureusing the following procedure: commercially available, cryopreservediPSCs were thawed using a water bath at 37° C. Cell suspension wasdiluted into commercially available media for stem cell proliferation(Miltenyi Biotec MACS iPS Brew) and seeded onto the ECM at approximately1,000 cells/cm² in a 6-well-plate with 2 mL of media/well. No Rockinhibitor was used. At day 1, the full volume of media was aspiratedgently from cells in culture and replaced with fresh media. Every 24hours, full media was replaced with fresh media. Once cells began toapproach confluence (as determined by brightfield microscopy), cellswere passage manually, using a sterile needle to cut large colonies intoapproximately 100 smaller colonies and then re-plate those by physicallylifting them off the dish with the sterile needle and placing them on afresh plate of the ECM. This procedure can be repeated indefinitely.

A photomicrograph showing Day 0 and Day 2 culture of iPSCs on amnioticfluid cell-derived ECM and a bone marrow cell-derived ECM is shown inFIG. 4.

A plot of iPSC colony growth curves of iPSCs cultured in the presence ofthe amniotic fluid cell-derived ECM and a bone marrow cell-derived ECMis shown in FIG. 5.

As can be seen in FIG. 4 and FIG. 5, the iPSCs proliferated in culturein the presence of the amniotic fluid cell-derived ECM, whereas iPSCscultured in the presence of a bone marrow cell-derived ECM had nogrowth.

Example 4—Preparation of a Cellular Construct of Mature Cardiomyocyteson an AFC-ECM and Maturation of Immature hiPSC-CMs on the AFC-ECM

Cellular constructs comprising monolayers of mature cardiomyocytes onextracellular matrices derived from cells derived in-vitro from amnioticfluid (AFC-ECMs) were prepared using the following method.

Using the methodology as outlined in Example 1, AFC-ECMs were preparedin 96-well plates (no silicone inserts were used). Using standard cellculture techniques, commercially available immature hiPSC-CMs fromCellular Dynamics International-FUJI (iCell® Cardiomyocytes) were platedon the AFC-ECMs. The immature hiPSC-CMs were plated at a density of50,000 cells per well (96 well plate) or 200,000 cells per well (6 wellplate), and were cultured for 7 days in RPMI media forming confluentmonolayers of mature cardiomyocytes on the AFC-ECMs, thereby formingcellular constructs of mature cardiomyocyte monolayers on the AFC-ECMs.

Over the 7-day period, the immature hiPSC-CMs were observed to matureinto the morphology and alignment of mature native adult cardiomyocytesas characterized by rod shaped cells with distinct sarcomere structure.For comparison purposes, iCell® immature hiPSC-CMs were also plated (nosilicone inserts were used) in 96-well plates and cultured in a similarfashion on standard Matrigel™ ECM, and on a bone marrow cell-derived ECM(BM-ECM) as prepared by methods as disclosed in U.S. Pat. No. 8,084,023,herein incorporated by reference. Results of the studies are shown inphotomicrographs of the cardiomyocytes on the different ECMs in FIGS. 6to 21.

As can be seen in the FIGS. 6 to 14, the cardiomyocytes on the AFC-ECMare mature, rod shaped cells with distinct sarcomere structureresembling native adult cardiomyocytes. The morphology and sarcomerestructure of the cardiomyocytes matured on AFC-ECM can be distinctlyseen by the presence of rod shaped cells with a striped appearanceidentified by arrows in the photomicrograph of FIG. 14. The figures showthe presence of fiber tracks on the AFC-ECM, and also show that themonolayer of mature cardiomyocytes is in alignment with the AFC-ECMwhich closely resembles characteristics found in native adultcardiomyocytes and native heart muscle tissue. By contrast, as can beseen in FIGS. 15 to 19 and FIGS. 20 to 21, the cardiomyocytes on thestandard Matrigel™ ECM and BM-ECM respectively, resemble fetal-likecardiomyocytes and do not have the characteristics of native adultcardiomyocytes or native heart muscle tissue.

Thus, the immature hiPSC-CMs cultured on the AFC-ECM achieved a higherstate of maturation than did the immature hiPSC-CMs cultured on thestandard Matrigel™ ECM or the natural cell-derived ECM from bone marrowcells. It is evident that the hiPSC-CM morphology was affecteddifferently by different ECM.

Example 5—High Throughput Cardiotoxicity Screen Testing of Drugs UsingCellular Construct of Mature Cardiomyocytes on an AFC-ECM

Cellular constructs of monolayers of mature cardiomyocytes on AFC-ECMwere prepared as described in Example 4 using 96-well plates (nosilicone inserts were used). After the 7-day maturation process theelectrophysiology of each well was observed using a plate reader usingthe following high throughput screen method. FluoVolt™ dye in HanksBalanced Salt Solution was loaded into each well. A high spatiotemporalCCD camera (SciMeasure DaVinci camera) combined with light emittingdiodes (LEDs) as shown in the schematic in FIG. 22. The camera and lenscombination were designed such that it allowed visualization of all thewells of the 96-well plate simultaneously with sufficient resolution toobserve action potential and calcium wave propagation. Each plate wascentered under the camera system, lighting was switched on and cameraacquisition was initiated and electrophysiological activity wasrecorded. Experiments were performed at about 37° C. Spontaneousactivity was recorded for at least 10 seconds and images were stored ona computer. After baseline readings were taken, 500 nM of the drug E4031(a hERG channel blocker) was added to each well. Images were analyzedand action potential duration, conduction velocity, beat rate andactivation patterns were quantified using image analysis software. Forcomparison purposes, immature hiPSC-CMs were cultured on standardMatrigel™ ECM and bone-marrow cell derived ECM (BM-ECM) in 96 wellplates (no silicone inserts were used) as described in Example 4, and500 nM of the drug E4031 (a hERG channel blocker) was analyzed insimilar fashion as with the AFC-ECM studies. Results of the E4031testing are shown in FIG. 23 and FIG. 24. As can be seen in FIG. 23, therecordings of the spontaneous action potentials recorded from thecardiomyocytes on the Matrigel™ ECM and BM-ECM showed that the drugE-4031 only caused action potential duration (APD) prolongation;however, on the mature cardiomyocytes on the AFC-ECM, the drug E-4031caused APD prolongation plus Rotors (TdP-like arrhythmias) which arearrhythmia activation patterns consistent with what is known to occur incases of TdP in humans. Thus, all three ECMs produced monolayers ofcardiomyocytes that responded with predicted APD prolongation, but onlythe AFC-ECM produced monolayers of cardiomyocytes that revealedtransition of APD prolongation to tachyarrhythmia, characteristic ofTdP. As can be seen in FIG. 24, 100% of the cardiomyocytes on theAFC-ECM responded to the drug E4031 with rotors (TdP like arrhythmia).

In additional studies, the drugs domperidone (3 μM), diisopyramide (100μM), azimilide (10 μM), D,1 Sotalol (100 μM), ibutilide (0.10 μM), andBepridil (10 μM) were tested with mature cardiomyocytes on AFC-ECM asprepared in Example 4 (no silicone inserts) in 96-well plates using theplate reader method described above. The results of the testing areshown in FIG. 25. As can be seen in the results, various types ofarrhythmias, i.e., tachyarrhythmia (TA), quiescence (Q), early afterdepolarization (EAD), were caused by the various drugs as notated in therecordings. In response to drugs classified as high risk for causing TdPfatal arrhythmias in patients by the FDA—these are the range ofarrhythmia types observed.

Various drugs shown in Table 3 and 4 below were also tested with maturecardiomyocytes on AFC-ECM as prepared in Example 4 (no silicone inserts)in 96-well plates using the plate reader method described above andobservations for any arrhythmia detected and APD90 prolongation at 10times the effective therapeutic plasma concentration (ETPC) werenotated. The drugs were selected from the CiPA Initiative's list ofcompounds for validation and testing of CiPA and are classified as highrisk, intermediate risk or low risk for causing fatal arrhythmias (TdPs)in patients. The complete list of CiPA compounds can be found athttp://cipaproject.org/wp-content/uploads/sites/24/2016/05/CiPA-Compounds.pdf.

TABLE 3 APD90 Risk Dose 1 Dose 2 Dose 3 Dose 4 Arrhythmia Prolongation?Drug Category (μM) (μM) (μM) (μM) Detected? @10X ETPC Tamoxifen Low 0.10.5 1.0 10.0 No No Nifedipine Low 0.001 0.01 0.1 1.0 Yes, Q No,Shortening Nitrendipine Low 0.00951 0.03004 0.09494 0.3 No No,Shortening Mexiletine Low 0.01 0.1 1.0 10.0 No No Ranolazine Low 0.010.1 1.0 10.0 No No Domperidone Intermediate 0.003 0.03 0.3 3.0 Yes, TAYes Droperidol Intermediate 0.03169 0.10014 0.31646 1.0 Yes, EAD YesClozapine Intermediate 0.09507 0.30043 0.94937 3.0 No Yes TerfenadineIntermediate 0.001 0.01 0.1 1.0 No Yes Disopyramide High 0.100 1.0010.00 100.00 Yes, EAD, TA Yes Quinidine High 0.95 3.00 9.49 30.0 Yes,EAD Yes D, I Sotalol High 0.1 1.0 10.0 100.00 Yes, EAD Yes Bepridil High0.01 0.10 1.00 10.0 Yes, Q Yes Dofetilide High 0.0005 0.0010 0.00320.010 Yes, EAD Yes

TABLE 4 APD90 Risk Dose 1 Dose 2 Dose 3 Dose 4 Arrhythmia Prolongation?Drug Category (μM) (μM) (μM) (μM) Detected? @10X ETPC Diltiazem Low 0.010.10 1.0 10.0 No No, shortening Loratadine Low 0.00095 0.003 0.009490.03 No No Metoprolol Low 3.169 10.0144 31.6456 100 Yes, No VerapamilLow 0.001 0.01 0.1 1 No No, Shortening Astemizole Intermediate 0.00010.001 0.01 0.1 Yes Yes Chlorpromazir Intermediate 0.09507 0.300430.94937 3.0 NA NA Cisapride Intermediate 0.00317 0.01001 0.03165 0.1 YesNo Pimozide Intermediate 0.00095 0.003 0.00949 0.03 Yes No RisperidoneIntermediate 0.00317 0.01001 0.03165 0.1 No Ondansetron Intermediate0.03 0.30 3.0 30.0 Yes, EAD Yes Azimilide High 0.01 0.10 1.0 10.0 Yes,EAD Yes Ibutilide High 0.0001 0.0010 0.0100 0.100 Yes, EAD YeSVandetanib High Yes, EAD Yes

Further analyses of the data from the drugs tested in Tables 3 and 4 areshown in FIGS. 26 to 29. FIG. 26 graphically shows the total number ofarrhythmias observed per any dose of each drug compound. FIG. 27graphically shows the relative occurrence of arrhythmias for each drugat specific clinically relevant doses, 10× the effective therapeuticplasma concentration (ETPC).

FIG. 28 graphically shows the action potential triangulation(APD90-APD30) in time (ms) for each drug. Triangulation is defined asthe repolarization time from APD30 to APD90. Action potentialtriangulation (APD90-APD30) is used as a predictor for proarrhythmia ofa drug. FIG. 29 shows the action potential triangulation in time (ms)for some drugs comparing cardiomyocyte assay performance of thecardiomyocytes on the AFC-ECM (SBS-AF Matrix) versus on Matrigel™ ECM.

FIG. 30 graphically shows the maximum drug-induced action potentialtriangulation of the listed drugs comparing cardiomyocyte performance ofiCell® hiPSC-CMs from Cellular Dynamics (blank circles) versus Cor.4U®hiPSC-CMs from Ncardia (dark circles) at any concentration of the listeddrugs. This figure is from publication Blinova et al, InternationalMultisite Study of Human-Induced Pluripotent Stem Cell-DerivedCardiomyocytes for Drug Proarrhythmic Potential Assessment, 2018, CellReports 24, 3582-3592. In contrast to FIG. 28, there is littlestratification between high risk and intermediate risk compounds in thedata set shown in FIG. 30. Thus, the AFC-ECM of this disclosure providesfor the production of more mature cardiomyocytes with more realisticfunction and drug responsiveness than of other natural cell-derived ECMsor Matrigel™ ECM.

Example 6—Observations of a Cellular Construct of Mature Cardiomyocyteson an AFC-ECM

Cellular constructs of mature cardiomyocytes on an AFC-ECM were preparedfollowing the procedures as described in Example 4 above with thefollowing modifications as noted below:

AFC-ECM was deposited onto Thermanox coverslips to enable immunostainingof cells and imaging using laser scanning confocal microscopy (NikonA1R).

Matrigel™ ECM was applied to a separate subset of Thermanox coverslipsfor comparison.

Cells from Cellular Dynamics International-FUJI (iCell® Cardiomyocytes)were plated as monolayers on these Thermanox coverslips coated with eachECM at a density of 200,000 cells per well in 6 well plates.

After 7 days of incubation in cell culture media, cells were fixed in 3%paraformaldehyde and processed for immunocytochemistry with applicationof commercially available primary antibodies to determine cellularexpression and localization in hiPSC-CMs. The following primaryantibodies for key cardiac myofilament proteins were used: Troponin I,α-actinin, cardiac troponin T (cTnT), cardiac troponin I (cTnI) andN-cadherin. Commercially available fluorescently labelled secondaryantibodies were used for detection. DAPI diamidino-2-phenylindole)fluorescent stain was used to mark the nuclei.

All procedures for cell labeling and visualization are as described inthe following references incorporated by reference herein. Herron, T. J.et al. Extracellular Matrix-Mediated Maturation of Human PluripotentStem Cell-Derived Cardiac Monolayer Structure and ElectrophysiologicalFunction. Circulation: Arrhythmia and Electrophysiology 9 (2016). daRocha, M. A. et al. Deficient cMyBP-C protein expression duringcardiomyocyte differentiation underlies human hypertrophiccardiomyopathy cellular phenotypes in disease specific human ES cellderived cardiomyocytes. J. Mol. Cell. Cardiol. 99, 197-206 (2016). daRocha, A. M. et al. hiPSC-CM Monolayer Maturation State Determines DrugResponsiveness in High Throughput Pro-Arrhythmia Screen. Sci. Rep. 7,13834 (2017).

Cell shape was quantified using fluorescent images analyzed in NISElements Software. Using cell area and perimeter, cellular circularitywas quantified using the established mathematical equation; CircularityIndex=4π*Area/Perimeter².

In some cases, mitochondria were stained using MitoTracker™ Red CMXRos(Thermo Fisher).

Results: Consistent with the results as seen in Example 4 above, datausing fluorescent labeling and imaging demonstrates that the AFC-ECMpromotes rapid (7-day) maturation of hiPSC-CMs, and shows that hiPSC-CMscultured on Matrigel™ ECM are circular in shape and have disorganizationof sarcomeres, whereas the same batch of hiPSC-CMs (isogenic coparator)cultured on AFC-ECM are rod shaped and have tightcompaction/organization of sarcomeres and myofilaments.

The photomicrophraphs in FIG. 31 show hiPSC-CMs cultured on Matrigel™ECM vs. AFC-ECM with immunofluorescent staining for Troponin I, usingDAPI to mark the nuclei. The photomicrophraphs in FIG. 32 show hiPSC-CMscultured on Matrigel™ ECM vs. AFC-ECM with immunofluorescent stainingfor α-actinin using DAPI to mark the nuclei. The photmicrographs in FIG.33 show hiPSC-CMs cultured on Matrigel™ ECM vs. AFC-ECM withimmunofluorescent staining for cTnT and N Cadherin, using DAPI to markthe nuclei. As can be seen in FIGS. 31 to 33, the hiPSC-CMs cultured onMatrigel™ ECM are circular in shape and have disorganization ofsarcomeres, whereas the same batch of hiPSC-CMs (isogenic coparator)cultured on AFC-ECM are rod shaped and have tightcompaction/organization of sarcomeres and myofilaments. Some examples ofthe cells are identified with arrows as either circular cells or rodshaped cells and some examples of the sarcomeres are identified witharrows in FIGS. 31 to 33. The photomicrographs in FIG. 34 show a singlehiPSC-CM cell cultured on Matrigel™ ECM vs. AFC-ECM withimmunofluorescent staining for cTnT using DAPI to mark the nuclei andshow that the cell cultured on Matrigel™ ECM is circular, whereas thecell cultured on AFC-ECM is rod shaped. The photomicrographs in FIG. 35show a single hiPSC-CM cell cultured on Matrigel™ ECM vs. AFC-ECM withimmunofluorescent staining for α-actinin using DAPI to mark the nucleiand show that the cell cultured on Matrigel™ ECM is circular, whereasthe cell cultured on AFC-ECM is rod shaped. The graph in FIG. 36 shows acomparison of the cellular circularity of the single cells shown in FIG.35 and shows a higher circularity index for the cell cultured on theMatrigel™ ECM indicating more roundness.

The photomicrographs in FIG. 37 show hiPSC-CMs cultured on Matrigel™ ECMvs. AFC-ECM with immunofluorescent staining for cTnI expression, usingDAPI to mark the nuclei. FIG. 38 shows western blotting of hiPSC-CMs onMatrigel™ ECM and AFC-ECM for cTnI expression and GAPDH (glyceraldehyde3-phosphate dehydrogenase). An analysis of the western blotting of FIG.38 is shown in the graph of FIG. 39 showing the cTnI expression relativeto GAPDH for the hiPSC-CMs on Matrigel™ ECM vs. AFC-ECM. The analysisshows a higher cTnI expression/GAPDH ratio for the hiPSC-CMs on theAFC-ECM than the hiPSC-CMs on the Matrigel™ ECM which indicates a morerobust cTnI expression from the hiPSC-CMs on the AFC-ECM than from thehiPSC-CMs on the Matrigel™ ECM.

The photomicrographs in FIG. 40 show hiPSC-CMs on Matrigel™ ECM vs.AFC-ECM stained for mitochondria with MitoTracker Red and show thatcells on AFC-ECM have more mitochondria and mitochondria with morepolarized inner membrane potential as evidenced by greater red signal.The graph in FIG. 41 shows the MitoTraker™ Red fluorescenceintensity/cardiomyocyte for hiPSC-CMs on Matrigel™ ECM vs. AFC-ECM andshows that the hiPSC-CMs on the AFC-ECM have higher MitoTraker™ Redfluorescence intensity indicating that these cells have moremitochondria and mitochondria with more polarized inner membranepotential than the hiPSC-CMs on the Matrigel™ ECM.

The photomicrographs (transmitted light) in FIG. 42 show hiPSC-CMscultured on Matrigel™ ECM and AFC-ECM that were coated on microelectrodearray (MEA) plates. As can be seen in FIG. 42, the hiPSC-CMs cultured onthe Matrigel™ ECM are circular in shape and the hiPSC-CMs cultured onthe AFC-ECM are rod shaped.

1. A method for the maturation of immature cardiomyocytes derived fromhuman induced pluripotent stem cells, the method comprising: (a)providing immature cardiomyocytes derived from human induced pluripotentstem cells (immature hiPSC-CMs); (b) providing an extracellular matrixderived in vitro from cells isolated from amniotic fluid (AFC-ECM); (c)contacting the immature hiPSC-CMs with the AFC-ECM; and (d) culturingthe immature hiPSC-CMs with the AFC-ECM in a culture media to inducematuration of the immature hiPSC-CMs, thereby forming maturecardiomyocytes; wherein the mature cardiomyocytes are characterized byrod shaped cells with distinct sarcomere structure resembling adulthuman cardiac tissue.
 2. The method of claim 1, wherein the immaturehiPSC-CMs are plated on the AFC-ECM.
 3. The method of claim 1, whereinthe mature cardiomyocytes form as a monolayer on the AFC-ECM, therebyforming a cellular construct comprising a monolayer of maturecardiomyocytes on the AFC-ECM, wherein the mature cardiomyocytes arealigned on the AFC-ECM.
 4. The method of claim 1, wherein the immaturehiPSC-CMs do not express inward-rectifier potassium channel Kir2.1.
 5. Acellular construct comprising a monolayer of mature cardiomyocytes on anextracellular matrix derived from cells isolated in vitro from amnioticfluid (AFC-ECM), wherein the mature cardiomyocytes are AFC-ECM culturedcardiomyocytes derived from human induced pluripotent stem cells(hiPSC-CMs), and wherein the mature cardiomyocytes are characterized byrod shaped cells with distinct sarcomere structure resembling adulthuman cardiac tissue.
 6. The cellular construct of claim 5, wherein themonolayer of mature cardiomyocytes is aligned on the AFC-ECM.
 7. Thecellular construct of claim 5, wherein fiber tracks are present on theconstruct.
 8. The cellular construct of claim 5, wherein the AFC-ECMcomprises laminin, collagen alpha-1 (XVIII), basement membrane-specificheparan sulfate proteoglycan core protein, agrin, vimentin, and collagenalpha-2 (IV), or isoforms thereof.
 9. The cellular construct of claim 8,wherein the isoform of collagen alpha-1 (XVIII) is isoform 2, or whereinthe isoform of agrin is isoform
 6. 10. The cellular construct of claim8, wherein the AFC-ECM further comprises fibronectin or an isoformthereof.
 11. The cellular construct of claim 5, wherein the AFC-ECM doesnot contain decorin, perlecan, or collagen (III).
 12. A method formaking a cellular construct of mature cardiomyocytes on an extracellularmatrix derived in vitro from cells isolated from amniotic fluid(AFC-ECM), the method comprising: (a) providing immature cardiomyocytesderived from human induced pluripotent stem cells (immature hiPSC-CMs),(b) providing an extracellular matrix derived in vitro from cellsisolated from amniotic fluid (AFC-ECM); (c) plating the immaturehiPSC-CMs on the AFC-ECM; (d) culturing the plated immature hiPSC-CMs onthe AFC-ECM in a culture media to induce maturation of the immaturehiPSC-CMs into mature cardiomyocytes and to form a monolayer of themature cardiomyocytes on the AFC-ECM, thereby forming the cellularconstruct; wherein the mature cardiomyocytes are characterized by rodshaped cells with distinct sarcomere structure resembling adult humancardiac tissue.
 13. The method of claim 12, wherein the monolayer ofmature cardiomyocytes is aligned on the AFC-ECM.
 14. The method of claim12, wherein fiber tracks are present on the cellular construct.
 15. Amethod for determining the cardiotoxicity or proarrhythmic effect of adrug compound in vitro, the method comprising contacting the drugcompound with the mature cardiomyocytes of any one of the cellularconstructs of claim 5, and observing for a change in theelectrophysiology of the mature cardiomyocytes to confirm whether thedrug compound has a cardiotoxic or proarrhythmic effect on the maturecardiomyocytes.
 16. The method of claim 15, wherein the change in theelectrophysiology of the mature cardiomyocytes is prolongation of actionpotential duration (APD), and wherein prolongation of APD confirms thatthe drug compound has a cardiotoxic or proarrhythmic effect on themature cardiomyocytes.
 17. The method of claim 15, wherein the change inthe electrophysiology of the mature cardiomyocytes is early afterdepolarization (EAD), and wherein early after depolarization (EAD)confirms that the drug compound has a cardiotoxic or proarrhythmiceffect on the mature cardiomyocytes.
 18. The method of claim 15, whereinthe change in the electrophysiology of the mature cardiomyocytes isdelayed after depolarization (DAD), and wherein delayed afterdepolarization (DAD) confirms that the drug compound has a cardiotoxicor proarrhythmic effect on the mature cardiomyocytes.
 19. The method ofclaim 15, wherein the change in the electrophysiology of the maturecardiomyocytes is action potential duration (APD) plus rotors, andwherein prolongation of APD plus rotors confirms that the drug compoundhas a cardiotoxic or proarrhythmic effect on the mature cardiomyocytes.20. The method of claim 15, wherein the change in the electrophysiologyof the mature cardiomyocytes is an arrhythmia, and wherein thearrhythmia confirms that the drug compound has a cardiotoxic orproarrhythmic effect on the mature cardiomyocytes.